Revision as of 11:12, 17 September 2015

FluID Detector

Lab Book

BioSpray

Our forensic toolkit aims to use synthetic biology approaches to improve on the current methods used by crime scene investigators.

Chromium Detector

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Fingerprint Aging

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Saliva

Week Beginning 1/6/2015

Summary

Lactoferrin binding protein A (LbpA) was successfully cloned into the vector pSB1C3.

1/6: Amplification of LbpA for Cloning into pSB1C3

Aim of Experiment: Amplify LbpA from MC58 N.meningitidis template chromosome.

Protocols Used: PCR

Results: Figure 1

Next Steps: Since the LbpA gene was amplified successfully, it was ready to be digested for subsequent cloning into pSB1C3.

2/6: Restriction Digests and Ligations with pSB1C3

Aim of experiment: To digest LbpA with BamHI and EcoRI and then ligate it into pSB1C3.

Protocols Used: Restriction Digests Ligations

Results: N/A

Next Steps: Ligations of pSB1C3-LbpA were left overnight until the next day when they would be transformed into the JM110 E.coli.

3/6: Transformations of pSB1C3-LpbA into JM110 E.coli

Aim of experiment: To transform the pSB1C3-LbpA ligations into JM110 E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: Transformations will be checked tomorrow and if successful, overnight cultures will be set up.

4/6: Overnight Cultures of JM110 E.coli containing pSB1C3-LbpA

Aim of experiment: To set up overnight cultures of JM110 E.coli containing pSB1C3-LbpA.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: A miniprep will be performed on the overnight cultures and a pre-sequence digest test performed tomorrow.

5/6: Plasmid Purification and Pre-Sequencing Digest Check

Aim of experiment: To miniprep the JM110 E.coli overnight cultures containing pSB1C3-LbpA.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with the highest concentration will be sent for sequencing.

Week Beginning 8/6/2015

Summary

Sequencing confirmed that LbpA had been successfully cloned into pSB1C3 therefore this week cloning was attempted to put LbpA into the high expression vector pQE80-L.

8/6: Amplification of LbpA for Cloning into pQE80-L

Aim of experiment: To amplify LbpA for subsequent cloninng into the pQE80-L vector.

Protocols Used: PCR

Results: Figure 2

Next Steps: The PCR product will be digested and ligated into pQE80-L tomorrrow.

9/6: Restriction Digests and Ligations of LbpAinto pQE80-L

Aim of experiment: To restrict and ligate the LbpA gene into the pQE80-L plasmid.

Protocols Used: Restriction Digests Ligations

Results:: N/A

Next Steps: Transformations of pQE80-L-LbpA into JM110 E.coli will be carried out tomorrow.

10/6: Transformations of pQE80-L-LpbA into JM110 E.coli

Aim of experiment: To transform pQE80-L-LpbA into JM110 E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: If the transformations are successful, colonies will be picked to set up overnight cultures.

Week Beginning 15/6/15

Summary

This week, cloning of LbpA into pQE80-L was continued due to last week's failed attempts.

15/6: PCR of LbpA for Cloning into pQE80-L

Aim of experiment: To re-amplify the LbpA gene for cloning into the pQE80-L vector.

Protocols Used: PCR

Results: Figure 3

Next Steps: The gel shows that LbpA has been successfully amplified so it will be digested and ligated into the pQE80-L vector tomorrow.

Week Beginning 22/6/15

Summary

LbpA was successfully cloned into the high expression vector pQE80-L.

23/6: Restriction Digest and Ligation of LbpA into pQE80-L

Aim of experiment: To digest the amplified LbpA gene with BamHI and KpnI and subsequently ligate it into the pQE80-L vector.

Protocols Used: Restriction Digests Ligations

Results: N/A

Next Steps: The ligations will be transformed into M15pREP4 E.coli.

24/6: Transformations of pQE80-L-LbpA into M15pREP4 E.coli

Aim of experiment: To transform pQE80-L-LbpA into M15pREP4 E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: Overnight cultures will be set up tomorrow if the transformations are successful.

25/6: Overnight Cultures of M15pREP4 E.coli containing pQE80-L-LbpA

Aim of experiment: To set up overnight cultures of M15pREP4 E.coli containing pQE80-L-LbpA.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: Overnight cultures will be miniprepped and a pre-sequence restriction digest will be performed to check for the presence of the LbpA gene.

26/6: Plasmid Purification of the M15pREP4 E.coli Overnight Cultures containing pQE80-L-LbpA

Aim of experiment: To miniprep the overnight cultures from yesterday and perform a pre-sequencing restriction digest using BamHI and KpnI to check for presence of LbpA gene.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: A sample of the miniprep was sent for sequencing to confirm the presence of LbpA in the pQE80-L vector.

Week Beginning 29/6/15

Summary

Protein expression optimization experiments were performed on LbpA to assay protein production levels. However, cell growth seems to cease when LbpA expression is induced.

1/7: M15pREP4 E.coli Overnight Cultures containing pQE80-L-LbpA

Aim of experiment: To set up overnight cultures in preparation for protein expression experiments tomorrow.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The overnight cultures will be used to set up day cultures tomorrow which will be used to test expression of LbpA.

2/7: Optimization of LbpA Expression

Aim of experiment: To set up day cultures and induce expression of LbpA using different concentrations of IPTG and check levels of protein production via SDS-PAGE.

Protocols Used: Protein Expression Optimization

Results: N/A

Next Steps: The samples were stored in the -20 ^oC freezer overnight and will be ran on a gel tomorrow.

3/7: Continuation of Optimization of LbpA Expression

Aim of experiment: To test the samples obtained yesterday on an SDS gel.

Protocols Used: Protein Expression Optimization

Results: Figure 4

Next Steps: Further optimization experiments will be set up next week using different concentrations of IPTG.

Week Beginning 6/7/15

Summary

Further protein expression optimization experiments were set up, this time using a range of IPTG concentrations to induce expression of LbpA.

7/7: Further Optimization of LbpA Expression

Aim of experiment: To set up further tests to optimize expression of LbpA using different concentrations of IPTG.

Protocols Used: Protein Expression Optimization Note: An SDS gel was not ran, instead the OD₆₀₀ readings were recorded for the uninduced and induced cultures at 20 minute intervals for 2 hours, 1 and half hours after the cultures were induced.

Results: Figure 5

Next Steps: A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after LbpA expression has been induced.

Week Beginning 13/7/15

Summary

Protein expression optimization experiments were continued this week. A growth curve assay was carried out to assess the effect of LbpA expression over a longer period of time.

14/7: Plate Reader Growth Curve Assay

Aim of experiment: To induce expression of LbpA using different concentration of IPTG and allow cells to grow for 16 hours in order to monitor cell growth.

Protocols Used: Growth Curve Assay

Results: Figure 6

Next Steps: A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after LbpA expression has been induced.

17/7: Sample Preparation for Western Blot

Aim of experiment: To check if the cells in the induced cultures ~7 hours after being induced are producing LbpA.

Protocols Used: Western Blot Sample Preparation Note: The day cultures were grown for 7 hours after they were induced and then the samples were frozen, to be blotted on Monday.

Results: N/A

Next Steps: The samples will be ran on an SDS gel on Monday and blotted.

Week Beginning 20/7/15

Summary

Samples from the growth curve assay were blotted to see if LbpA is being produced at the later stages of the growth curve. A further growth curve assay was also performed which monitored cell growth until a certain OD₆₀₀ was reached, at which point the cultures were induced.

20/7: Western Blot LpbA Culture Samples

Aim of experiment: To western blot the LbpA culture samples from Friday.

Protocols Used: Western Blot

Results: Nothing was visible on the blot.

Next Steps: Another plate reader experiment will be set up on Thursday where the cultures will be monitored for three hours before induction with IPTG and then for a further 20 hours afterwards.

23/7: Growth Curve Assay

Aim of experiment: To check if the cells begin to die after induction with IPTG after having already grown to an OD₆₀₀ of 0.5.

Protocols Used: Growth Curve Assay

Results: Figure 7

Next Steps: Samples of these cultures will be taken 6 hours after induction to check for the presence of LbpA.

Week Beginning 27/7/15

Summary

The presence of LbpA was confirmed in the culture 6 hours after induction with IPTG so protein purification was started this week.

27/7: Overnight Cultures of M15pREP4 E.coli containing pQE80-L-LbpA

Aim of experiment: To set up a 150 ml overnight culture for sub-culturing tomorrow.

Protocols Used: Overnight Cultures Note: 150ml of LB was used and 150µl of each antibiotic.

Results: N/A

Next Steps:150ml of fresh LB will be inoculated tomorrow with 7.5 ml of the overnight cultures.

28/7: LbpA Expression Assay

Aim of experiment: To set up day cultures using the overnight cultures from yesterday and test for expression of LbpA 6 hours after induction.

Protocols Used: Protein Expression Test Note: 150 ml of fresh LB was inoculated with 7.5 ml of the overnight cultures. Only an uninduced control and an 1 mM IPTG induced culture was set up. The samples were not blotted, they were only ran on an SDS gel.

Results: Figure 8

Next Steps: A 3 L culture will be set up on Thursday under the same conditions as the overnight cultures in order to try and purify LbpA.

29/7: Overnight Cultures of M15pREP4 E.coli containing pQE80-L-LbpA for 3L Culture

Aim of experiment: To set up overnight cultures for the 3L day cultures that will be grown tomorrow.

Protocols Used: Overnight Cultures Note: One 150 ml overnight culture was set up using 150 µl of each antibiotic.

Results: N/A

Next Steps: The 3 L culture will be set up tomorrow allowed to grow to OD₆₀₀ of 0.6-1 and then induced with 1 mM IPTG and grown for 6 hours.

30/7: 3 L Culture of M15pREP4 E.coli containing pQE80-L-LbpA for Purification of LbpA

Aim of experiment: To set up 3 L day cultures for purifying LbpA.

Protocols Used: Protein Purification Note: The procedure was stopped at Step 6 and the pellets were frozen at -20 ^oC.

Results: N/A

Next Steps: Protein purification will be continued on Monday.

Week Beginning 3/8/15

Summary

The first round of LbpA purification failed at the affinity chromatography stage so another 6 L culture was grown this week to attempt purification again.

3/8: Continuation of LbpA Purification

Aim of experiment: To continue with purification of LbpA.

Protocols Used: Protein Purification Note: The protocol was continued from Step 7

Results: N/A

Next Steps:Size exclusion chromatography was not carried out since an SDS gel showed LbpA was not present after performing affinity chromatography. Another 6L culture will be set up this week.

5/8: Overnight Cultures of M15pREP4 E.coli containing pQE80-L-LbpA

Aim of experiment: To set up overnight cultures for the 6 L culture that will be set up tomorrow.

Protocols Used: Overnight Cultures Note: One 150 ml overnight culture was set up using 150 µl of each antibiotic.

Results: N/A

Next Steps:The overnight cultures will be used to set up the 6 L day culture tomorrow.

6/8: 6 L Day Cultures of M15pREP4 E.coli containing pQE80-L-LbpA

Aim of experiment: To set up 6 L day cultures for purifying LbpA and begin the purification process.

Protocols Used: Protein Purification Note: The protocol was stopped at Step 6.

Results: N/A

Next Steps:Purification of LbpA will be continued on Monday.

Week Beginning 10/8/15

Summary

Purification of LbpA was continued but unfortunately was unsuccessful. If time permits, another round of purification may be reattempted.

10/8: Continuation of LbpA Purification

Aim of experiment: To continue with purification of LbpA.

Protocols Used: Protein Purification Note: The protocol was continued from Step 7

Results: N/A

Next Steps: Purification of LbpA will be continued tomorrow.

11/8: Continuation of LbpA Purification

Aim of experiment: To purify LbpA using affinity chromatography.

Protocols Used: Protein Purification Note: The protocol was stopped at Step 13.

Results: N/A

Next Steps:Since no peaks were observed, purification of LbpA was abadoned at this stage. Purification may be retried.

Semen

Week Beginning 25/5/2015

Summary

Cloning of PotD into the biobrick vector, pSB1C3, was started this week.

28/5: PCR of PotD from MG1655 E.coli gDNA

Aim of Experiment: To amplify PotD from MG1655 E.coli gDNA.

Protocols Used: PCR

Results: N/A

Next Steps: The PCR product will be gel extracted, digested and ligated into the biobrick vector pSB1C3 tomorrow.

29/5: Gel Extraction, Restriction Digest and Ligation of SBP into pSB1C3

Aim of experiment: To gel extract the PotD PCR product and digest it with PstI and EcoRI. A ligation will then be set up with pSB1C3.

Protocols Used: Restriction Digest Ligation

Results: N/A

Next Steps: Ligations will be transformed into JM110 E.coli on Monday.

Week Beginning 1/6/2015

Summary

Cloning of PotD into the biobrick vector was completed this week.

1/6: Transformation of pSB1C3-PotD into JM110 E.coli

Aim of experiment: To transform pSB1C3-PotD into JM110 E.coli.

Protocols Used: Transformations

Results:N/A

Next Steps: If the transformations are successful, overnight cultures will be set up tomorrow.

2/6: Overnight Cultures of pSB1C3-PotD in JM110 E.coli

Aim of experiment: To set up overnight cultures of pSB1C3-PotD in JM110 E.coli

Protocols Used: Overnight Cultures

Results:: N/A

Next Steps: The overnight cultures will be miniprepped tomorrow.

3/6: Plasmid Purification of pSB1C3-PotD from Overnight Cultures

Aim of experiment: To miniprep the overnight cultures from yesterday.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with the highest concentration will be sent for sequencing to confirm PotD has been inserted into pSB1C3 successfully.

Week Beginning 15/6/15

Summary

This week, cloning of PotD into pQE80-L was started. The gene for Spermine Binding Protein (SBP) also arrived in a plasmid so it was transformed into MC1061 E.coli and plasmid purified.

15/6: PCR of PotD for Cloning into pQE80-L

Aim of experiment: To amplify the PotD gene for cloning into the pQE80-L vector.

Protocols Used: PCR

Results: Figure 3

Next Steps: The gel shows that PotD has been successfully amplified so it will be digested and ligated into the pQE80-L vector tomorrow.

16/6: Gel Extraction of PotD PCR Product

Aim of experiment: To gel extract the PotD PCR product.

Protocols Used: PCR

Results: Figure 3

Next Steps: The gel shows that PotD has been successfully amplified so it will be digested and ligated into the pQE80-L vector tomorrow.

17/6: Restriction Digest and Ligation of PotD into pQE80-L. Transformation of IDT Plasmid containing SBP into MC1061 E.coli

Aim of experiment: To digest the PotD PCR product with KpnI and BamHI and then ligate it into pQE80-L. To transform MC1061 E.coli with the IDT plasmid containing SBP.

Protocols Used:

PotD: Restriction Digests Ligations SBP:Transformations

Results: N/A

Next Steps: The pQE80-L-PotD ligations will be transformed into MC1061 E.coli. Overnight cultures of the transformations will be set up tomorrow.

18/6: Transformations of PotD + pQE80-L into MC1061 E.coli. Overnight Cultures of MC1061 E.coli with SBP + IDT Plasmid

Aim of experiment: To transform the pQE80-L-PotD ligations into MC1061 E.coli. To set up overnight cultures of the transformations of MC1061 E.coli with SBP + IDT Plasmid that were set up yesterday.

Protocols Used: PotD: Transformations SBP: Overnight Cultures

Results:N/A

Next Steps: If transformations are successful, overnight cultures will be set up tomorrow.

19/6: Repeat of Ligation of PotD into pQE80-L. Plasmid Purification of the IDT Plasmid Containing SBP

Aim of experiment: To repeat ligations of PotD into pQE80-L since transformations did not work. To miniprep the overnight cultures that were set up yesterday.

Protocols Used: PotD: Ligations SBP: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: Ligations will be left over the weekend and transformed on Monday.

Week Beginning 22/6/15

Summary

Cloning of PotD into the high expression vector pQE80-L was completed. Cloning of SBP into pSB1C3 and pQE80-L was started this week.

22/6: Transformation of pQE80-L-PotD Ligations. PCR and Gel Extraction of SBP for Cloning into pQE80-L and Restriction Digests and Ligation of SBP for Cloning into pSB1C3

Aim of Experiment: To transform pQE80-L-PotD into MC1061E.coli. To set up a PCR of SBP using the purified plasmid from Friday and to gel extract the PCR product for cloning into pQE80-L. To digest SBP out of the same plasmid using EcoRI and PstI and subsequently ligate it into pSB1C3.

Protocols Used: PotD: Transformations SBP: PCR Restriction Digests Ligations

Results: SBP : Figure 1

Next Steps: Overnight cultures will be set up tomorrow for the PotD transformations. The SBP PCR product will be digested and ligated into pQE80-L tomorrow. The pSB1C3-SBP ligations will be transformed into MC1061 E.coli.

23/6: Overnight Cultures of MC1061 E.coli containing pSB1C3-PotD. Transformations of MC1061 E.coli with pSB1C3 + SBP. Restriction Digests and Ligations of SBP into pQE80-L and Transformations into MC1061 E.coli

Aim of experiment: To set up overnight cultures of MC1061 E.coli containing pSB1C3 + PotD. To transform MC1061 E.coli with pSB1C3 + SBP. To digest the SBP PCR product with BamHI and Kpn I, ligate it into pQE80-L and transform these ligations into MC1061 E.coli.

Protocols Used: PotD: Overnight Cultures SBP: Transformations Restriction Digests Ligations

Results: N/A

Next Steps: The MC1061 E.coli overnight cultures containing pQE80-L + PotD will be miniiprepped tomorrow. Overnight cultures will be set up of the MC1061 E.coli transformations with pSB1C3 + SBP and pQE80-L + SBP.

24/6: Plasmid Purification of the Overnight Cultures of MC1061 E.coli containing pSB1C3-PotD. Overnight Cultures of the Transformations of MC1061 E.coli with pSB1C3 + SBP and pQE80-L + SBP.

Aim of experiment: To miniprep the overnight cultures of MC1061 E.coli containing pSB1C3-PotD. To set up overnight cultures of the transformations of MC1061 E.coli with pSB1C3-SBP and pQE80-L-SBP

Protocols Used: PotD: Plasmid Purification (QIAprep® Spin Miniprep Kit) SBP: Overnight Cultures

Results: N/A

Next Steps: The miniprep with the highest concentration for pQE80-L + PotD was sent for sequencing. The SBP overnight cultures will be miniprepped tomorrow.

25/6: Plasmid Purification of the Overnight Cultures of MC1061 E.coli containing pSB1C3-SBP and pQE80-L-SBP

Aim of experiment: To miniprep the overnight cultures of MC1061 E.coli containing pSB1C3-SBP and pQE80-L + SBP.

Protocols Used: SBP: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with the highest concentration for both pSB1C3-SBP and pQE80-L-SBP were sent for sequencing.

Week Beginning 29/6/15

Summary

Sequencing showed SBP failed to insert successfully into pSB1C3 so cloning was repeated from the ligation stage this week. However, SBP and PotD were successfully inserted into the high expression vector pQE80-L, so overexpression assays were started this week for both.

29/6: Ligations of SBP into pSB1C3 and Transformations into MC1061 E.coli

Aim of Experiment: To ligate SBP into pSB1C3 and transform the ligations into MC1061 E.coli.

Protocols Used: SBP: Ligations Transformations

Results: SBP : Figure 1

Next Steps: Overnight cultures will be set up tomorrow if transformations are successful.

30/6: Overnight Cultures of MC1061 E.coli containing pSB1C3 + SBP, pQE80-L + SBP and pQE80-L + PotD

Aim of experiment: To set up overnight cultures of MC1061 E.coli containing pSB1C3 + SBP. To set up overnight cultures of pQE80-L + SBP and pQE80-L + PotD in MC1061 E.coli for overexpression assays to be performed tomorrow.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The MC1061 E.coli overnight cultures containing pSB1C3 + SBP will be miniiprepped tomorrow. The MC1061 E.coli overnight cultures containing pQE80-L + SBP and pQE80-L + PotD will be subcultured tomorrow in order to perform overexpression assays.

1/7: Plasmid Purification of the Overnight Cultures of MC1061 E.coli containing pSB1C3 + SBP. PotD and SBP Expression Optimization

Aim of experiment: To miniprep the overnight cultures of MC1061 E.coli containing pSB1C3 + SBP. To start protein expression optimization for PotD and SBP.

Protocols Used: SBP: Plasmid Purification (QIAprep® Spin Miniprep Kit) SBP and PotD: Protein Expression Optimization Note: Protocol was stopped at Step 8. Samples will be ran on an SDS gel tomorrow.

Results: N/A

Next Steps: The miniprep with the highest concentration for pSB1C3 + SBP was sent for sequencing. The PotD and SBP samples will be ran on an SDS gel tomorrow.

2/7: Continuation of PotD and SBP Expression Optimization

Aim of experiment: To run the samples of PotD and SBP on an SDS gel and begin Western Blotting.

Protocols Used: PotD and SBP: Protein Expression Optimization Note: Protocol was continued from Step 9.

Results: Figure 2

Next Steps: The membrane for the Western blot has been left at the blocking stage so will be continued tomorrow.

3/7: Western Blot of PotD and SBP Day Culture Samples

Aim of experiment: To continue the Western Blot of PotD and SBP.

Protocols Used: PotD and SBP: Western Blot

Results: Figure 3

Next Steps: PotD has been successfully characterized so purification of this protein will be started next week. Further expression optimization experiments for SBP will be set up next week using different concentrations of IPTG, since its characterization was unsuccessful.

Week Beginning 6/7/15

Summary

Further protein expression optimization experiments were set up, this time using a range of IPTG concentrations to induce expression of SBP.

6/7: Further Optimization of SBP Expression

Aim of experiment: To set up further tests to optimize expression of SBP using different concentrations of IPTG.

Protocols Used: Protein Expression Optimization Note: Cultures were induced with o.5mM, 1mM and 2mM IPTG. The Western Blot was left to block overnight.

Results: Figure 4

Next Steps: The Western Blot will be completed tomorrow.

7/7: Western Blot of SBP

Aim of experiment: To complete the Western Blot that was started yesterday.

Protocols Used: Western Blot Note: Protocol was continued from Step 12.

Results: Nothing was visible on the blot.

Next Steps: A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after SBP expression has been induced.

Week Beginning 13/7/15

Summary

Protein expression optimization experiments were continued this week. A growth curve assay was carried out to assess the effect of SBP expression over a longer period of time. Purification of PotD was also started this week.

14/7: Plate Reader Growth Curve Assay

Aim of experiment: To induce expression of SBP using different concentration of IPTG and allow cells to grow for 16 hours in order to monitor cell growth.

Protocols Used: Growth Curve Assay

Results: Figure 5

Next Steps: A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after SBP expression has been induced.

15/7: Overnight Cultures for 3L Culture for PotD Purification

Aim of experiment: To set up overnight cultures that will be used tomorrow to set up 3L cultures for PotD purification.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The 3L day culture will be set up tomorrow.

16/7: Sample Preparation for Western Blot of SBP and 3L Day Culture for PotD Purification

Aim of experiment: To check if the cells in the induced cultures ~7 hours after being induced are producing SBP. To set up 3L Day Culture for PotD Purification

Protocols Used: SBP: Western Blot Sample Preparation Note: The day cultures were grown for 7 hours after they were induced and then the samples were frozen, to be blotted on Monday. PotD: Protein Purification Note: Cultures were left to grow overnight after being induced.

Results: N/A

Next Steps: The samples will be ran on an SDS gel on Monday and blotted. The PotD cultures will be harvested tomorrow.

17/7: Harvesting PotD Cultures

Aim of experiment: To harvest the day cultures grown yesterday for PotD purification.

Protocols Used: Protein Purification Note: After the cultures were spun down the pellets were frozen.

Results: N/A

Next Steps: Purification of PotD will continue on Monday.

Week Beginning 20/7/15

Summary

Samples from the growth curve assay were blotted to see if SBP is being produced at the later stages of the growth curve. A further growth curve assay was also performed which monitored cell growth until a certain OD₆₀₀ was reached, at which point the cultures were induced. Purification of PotD was also continued this week.

20/7: Western Blot SBP Culture Samples

Aim of experiment: To western blot the SBP culture samples from Friday. To continue purfying PotD.

Protocols Used: SBP: Western Blot PotD: Protein Purification Note: Affinity chromatography was completed but no peaks were observable.

Results: Nothing was visible on the blot.

Next Steps: Another plate reader experiment will be set up on Thursday where the cultures will be monitored for three hours before induction with IPTG and then for a further 20 hours afterwards. Another 150ml overnight culture was set up for PotD so 3L cultures could be set up tomorrow.

21/7: 3L Day Cultures for PotD Purification

Aim of experiment: To set up 3L Day Culture for PotD Purification.

Protocols Used: Protein Purification Note: Cultures were left to grow overnight after being induced.

Results: N/A

Next Steps: Purification will be continued tomorrow.

22/7: Continuation of PotD Purification

Aim of experiment: To continue purification of PotD.

Protocols Used: Protein Purification Note: The protocol was continued from Step 4 and stopped at Step 13.

Results: N/A

Next Steps: Purification will be continued tomorrow.

23/7: Continuation of PotD Purification and Growth Curve Assay for SBP

Aim of experiment: To continue purification of PotD and to check if the cells begin to die after inducing expression of SBP with IPTG after having already grown to an OD₆₀₀ of 0.5.

Protocols Used: PotD: Protein Purification Note: The protocol was continued from Step 13. SBP: Growth Curve Assay

Results: Figure 6

Next Steps:The samples from the size exclusion chromatography performed on PotD will be prepared tomorrow for blotting next week. For SBP further overexpression tests will be set up next week.

24/7: SDS-PAGE and Western Blot of PotD

Aim of experiment: To run SDS-PAGE of the elutions obtained on Friday after SEC.

Protocols Used: SDS-PAGE

Results: Figure 7

Next Steps:The same samples will be Western Blotted on Monday to confirm that the bands oberved are PotD.

Week Beginning 27/7/15

Summary

PotD was successfully purified and characterized this week. SBP was still unobservable in induced 150ml cultures so 3L cultures containing pQE80-L-SBP were set up.

27/7: Overnight Cultures for SBP Expression Assays and SDS-PAGE and Western Blot of PotD

Aim of experiment: To set up a 150ml overnight culture containing pQE80-L-SBP for sub-culturing tomorrow and to run an SDS-PAGE and Western Blot the PotD samples from Friday.

Protocols Used: SBP: Overnight Cultures Note: 150ml of LB was used and 150µl of each antibiotic. PotD: SDS-PAGE and Western Blotting Note: The membrane was left blocking overnight.

Results: N/A

Next Steps:150ml of fresh LB will be inoculated tomorrow with 7.5ml of the overnight cultures containing pQE80-L-SBP. The PotD Western Blot will be completed tomorrow.

28/7: SBP Expression Assay and Completion of PotD Westen Blot

Aim of experiment: To set up day cultures using the overnight cultures from yesterday and test for expression of LbpA 6 hours after induction.

Protocols Used: Protein Expression Test Note: 150ml of fresh LB was innoculated with 7.5ml of the overnight cultures. Only an uninduced control and an 1mM IPTG induced culture was set up. The samples were not blotted, they were only ran on an SDS gel.

Results: SBP: Nothing was observable on the SDS gel. PotD: Figure 8

Next Steps: Since PotD has now been successful purified, the next step is to attach a fluorescent nanobead to the protein. Hopefully, this will be attempted in the coming weeks. Even though SBP was not visible on the SDS gel, 3L cultures will be set up this week to try and harvest the protein since the culture size may be too small.

29/7: Overnight Cultures for 3L Culture

Aim of experiment: To set up overnight cultures containing pQE80-L-SBP for the 3L day cultures that will be grown tomorrow.

Protocols Used: Overnight Cultures Note: One 150ml overnight culture was set up using 150µl of each antibiotic.

Results: N/A

Next Steps: The 3L culture will be set up tomorrow allowed to grow to OD₆₀₀ of 0.6-1 and then SBP expression induced with 1mM IPTG and grown for 6 hours.

30/7: 3L Culture for Purification of SBP

Aim of experiment: To set up 3L day cultures for purifying SBP.

Protocols Used: Protein Purification Note: The procedure was stopped at Step 6 and the pellets were frozen at -20^oC.

Results: N/A

Next Steps: Protein purification will be continued on Monday.

Week Beginning 3/8/15

Summary

It seems that SBP has been successfully purified and characterized. However, there was still a lot of contamination in the samples after size eclusion chromatography was performed. .

3/8: Continuation of SBP Purification

Aim of experiment: To continue with purification of SBP.

Protocols Used: Protein Purification Note: The protocol was continued from Step 7

Results: N/A

Next Steps:Size exclusion chromatography will be carried out tomorrow.

4/8: Continuation of SBP Purification

Aim of experiment: To continue purification of SBP.

Protocols Used: Protein Purification Note: The protocol was continued from Step 14.

Results: N/A

Next Steps: The fractions corresponding to the peak observed during SEC, will be ran on an SDS gel and Western Blotted tomorrow.

6/8: Western Blot of SBP

Aim of experiment: To perform a Western Blot on the samples obtained from Size Exclusion Chromatography yesterday..

Protocols Used: Western Blot

Results: Figure 9

Next Steps: There was still some contamination in the gel filtrated samples so this will have to be removed before a nanobead can be attached to the protein.

Week Beginning 10/8/15

Summary

Interactions between PotD and spermidine were analysed via tryptophan fluorescence this week.

10/8: Tryptophan Fluorescence

Aim of experiment: To analyse the interaction of PotD and spermidine using tryptophan fluorescence.

Protocols Used: Tryptophan Fluorescence

Results: N/A

Next Steps:

Blood

Week Beginning 10/6/2015

Summary

Cloning of hHBA and hHBB into the biobrick vector, pSB1C3, was performed this week.

10/6: Restriction Digests of hHBA and hHBB from IDT gBlocks and Ligation into pSB1C3

Aim of Experiment: To digest hHBA and hHBB from IDT gBlocks using EcoRI and PstI for ligation into pSB1C3.

Protocols Used: Restriction Digests Ligations

Results: N/A

Next Steps: The pSB1C3-hHBA and pSB1C3-hHBB ligations will be transformed into JM110 E.coli.

11/6: Transformations of pSB1C3-hHBA and pSB1C3-hHBB Ligations into JM110 E.coli

Aim of experiment: To transform the pSB1C3-hHBA and pSB1C3-hHBB ligations into JM1110 E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: If transformations are successful, overnight cultures will be set up.

Week Beginning 15/6/2015

Summary

The sequence for pSB1C3-hHBB was confirmed this week so cloning of hHBB into the two hybrid system vector pUT18 was started. Sequencing of pSB1C3-hHBA was incorrect so another miniprep was sent for sequencing.

15/6: Overnight Cultures of JM110 E.coli containing pSB1C3-hHBA and pSB1C3-hHBB

Aim of experiment: To set up overnight cultures of JM110 E.coli containing pSB1C3-hHBA and pSB1C3-hHBB from the transformations performed on Tuesday.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: Overnight cultures will be miniprepped tomorrow and a test restriction digest will be performed to check for the presence of the inserts.

16/6: Plasmid Purification of the JM110 E.coli Overnight Cultures containing pSB1C3-hHBA and pSB1C3-hHBB

Aim of experiment: To miniprep the JM110 E.coli overnight cultures containing pSB1C3-hHBA and pSB1C3-hHBB and perform a pre-sequence digest check.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit) Restriction Digests

Results: Figure 1

Next Steps: The gel indicates that hHBA and hHBB have inserted successfully into pSB1C3 so samples of the minipreps will be sent for sequencing to confirm this.

17/6: Amplification of hHBA and hHBB for Cloning into the Two Hybrid System Vectors

Aim of experiment: To amplify hHBA and hHBB for Cloning into the Two Hybrid System Vectors pT25 and pUT18, respectively.

Protocols Used: PCR

Results: Figure 2
Next Steps: Sequencing of pSB1c3-hHBA and pSB1c3-hHBB showed that pSB1c3-hHBA had a 1bp deletion so one of the other minipreps was sent for sequencing. pSB1c3-hHBB, however, was successful so tHe band for hHBB will be gel extracted and digested tomorrow.

18/6: Gel Extraction and Restriction Digests hHBB for Cloning into pUT18

Aim of experiment: To gel extract and digest hHBB with BamHI and EcoRI.

Protocols Used: Restriction Digests

Results: N/A
Next Steps: Ligations of hHBB will be set up tomorrow.

19/6: Ligations of hHBB into pUT18

Aim of experiment: To set up ligations of pUT18-hHBB.

Protocols Used: Ligations

Results: N/A
Next Steps: Ligations will be left over the weekend and transformed into BTH101 E.coli.

Week Beginning 22/6/15

Summary

Cloning of hHBA and hHBB into pT25 and pUT18, respectively, was continued this week.

22/6: Transformations of pUT18-hHBB into BTH101 E.coli

Aim of experiment: To transform pUT18-hHBB into BTH101 E.coli.

Protocols Used: Transformations

Results:N/A

Next Steps:If transformations are successful, overnight cultures will be set up tomorrow. Sequencing showed the sequence for pSB1C3-hHBA was incorrect again, a third miniprep will be sent for sequencing.

23/6: Overnight Cultures of BTH101 E.coli containing pUT18-hHBB

Aim of experiment: To set up overnight cultures of BTH101 E.coli containing pUT18-hHBB from the transformations carried out yesterday.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The overnight cultures will be miniprepped and sent for sequencing tomorrow.

24/6: Plasmid Purification of the BTH101 E.coli Overnight Cultures containing pUT18-hHBB

Aim of experiment: To miniprep the BTH101 E.coli Overnight Cultures containing pUT18-hHBB.

Protocols Used:

Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with the highest concentration will be sent for sequencing to confirm presence of insert. Sequencing for pSB1C3-hHBA came back correct so another PCR of hHBA will be set up tomorrow for cloning into pT25.

25/6: Amplification of hHBA for Cloning into pT25

Aim of experiment: To amplify hHBA for Cloning into pT25

Protocols Used: PCR

Results: Figure 3

Next Steps: The PCR product digested and ligated into pT25 tomorrow.

26/6: Restriction Digests and Ligations of hHBA for Cloning into pT25

Aim of experiment: To digest the hHBA PCR product with BamHI and KpnI and set up ligations with pT25.

Protocols Used: Restriction Digests Ligations

Results: N/A

Next Steps: Ligations will be left over the weekend and transformed into BTH101 E.coli on Monday.

Week Beginning 29/6/15

Summary

Cloning of hHBA into pT25 was completed this week. Sequencing confirmed that hHBB has been successfully inserted into pUT18. Cloning of both hHBA and hHBB, individually, into the high expression vector pQE80-L was started this week.

29/6: Transformation of pT25-hHBA Ligations into BTH101 E.coli/span>

Aim of Experiment: To transform pT25-hHBA into BTH101 E.coli.

Protocols Used: Transformations>

Results: N/A

Next Steps: Overnight cultures will be set up tomorrow for the pT25-hHBA transformations if successful. Sequencing confirmed hHBB has been successfully inserted into pUT18.

30/6: Overnight Cultures of MC1061 E.coli containing pUT18-hHBA and Amplification of hHBA and hHBB for Cloning into pQE80-L

Aim of experiment: To set up overnight cultures of MC1061 E.coli containing pT25-hHBA. To amplify hHBA.and hHBB for cloning into pQE80-L.

Protocols Used: Overnight Cultures Transformations Restriction Digests PCR

Results: Figure 4

Next Steps: The BTH101 E.coli overnight cultures containing pT25-hHBB will be miniiprepped tomorrow. The PCR products will be digested and ligate dinto pQE80-L tomorrow.

1/7: Plasmid Purification of the Overnight Cultures of BTH101 E.coli containing pT25-hHBA and Restriction Digests and Ligations of hHBA and hHBB into pQE80-L

Aim of experiment: To miniprep the overnight cultures of BTH101 E.coli containing pT25-hHBA. To digest hHBA and hHBB with BamHI and KpnI and set up ligations with pQE80-L.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit) Restriction Digests Ligations

Results: N/A

Next Steps: The miniprep with the highest concentration for pT25-hHBA will be sent for sequencing. The pQE80-L-hHBA and pQE80-L-hHBB will be transformed into M15[pREP4] E.coli tomrorrow.

2/7: Transformations of pQE80-L-hHBA and pQE80-L-hHBB Ligations into M15[pREP4] E.coli

Aim of experiment: To transform the pQE80-L-hHBA and pQE80-L-hHBB ligations into M15[pREP4] E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: If transformations are successful, overnight cultures will be set up on Monday.

Week Beginning 6/7/15

Summary

Cloning of hHBA and hHBB into pQE80-L was completed this week. Cloning of Haptoglobin (hHBN) was also started this week.

6/7: Overnight Cultures of m15[pREP4] E.coli containing pQE80-L-hHBA and pQE80-L-hHBB

Aim of Experiment: To set up overnight cultures of BTH101 E.coli containing pQE80-L-hHBA and pQE80-L-hHBB.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps:The overnight cultures will be miniprepped tomorrow. Sequencing for pT25-hHBA was incorrect so cloning will be repeated from the restriction digest stage.

7/7: Plasmid Purification of m15[pREP4] E.coli Overnight Cultures containing pQE80-L-hHBA and pQE80-L-hHBB

Aim of experiment: To set up miniprep the BTH101 E.coli Overnight Cultures containing pQE80-L-hHBA and pQE80-L-hHBB.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: Figure 5

Next Steps: The pre-sequence digest indicates that both hHBA and hHBB have been inserted into pQE80-L successfully. The minipreps with the highest concentration will be sent for sequencing. The sequence for Haptoglobin (hHBN) arrived in an IDT plasmid, work will be be started on this tomorrow.

8/7: Transformation of IDT Plasmid-hHBN into MC1061 E.coli

Aim of experiment: To transform the IDT Plasmid-hHBN into MC1061 E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: Overnight cultures will be set up tomorrow.

9/7: Overnight Cultures of MC1061 E.coli containing IDT Plasmid-hHBN

Aim of experiment: To set up overnight cultures of MC1061 E.coli containing IDT Plasmid-hHBN.

Results: N/A

Next Steps: The overnight cultures will be miniprepped tomorrow and hHBN will be amplified for cloning into pSB1C3.

10/7: Plasmid Purification of IDT Plasmid-hHBN and Digestion of hHBN for Cloning into pSB1C3

Aim of experiment: To miniprep the MC1061 E.coli containing IDT Plasmid-hHBN and to digest hHBN out of the plasmid using EcoRI and PstI.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit) Restriction Digests

Results: N/A

Next Steps: hHBN will be ligated into pSB1C3 on Monday. Sequencing also confirmed hHBA and hHBB have been successfully inserted into pQE80-L.

Week Beginning 13/7/15

Summary

hHBB was successfully characterized this week so protein purification was started. Cloning of hHBN into pSB1C3 was also completed.

13/7: Ligations of hHBN into pSB1C3 and Overnight Cultures of M15[pREP4] E.coli containing pQE80-L-hHBA and pQE80-L-hHBB

Aim of experiment: To ligate hHBN into pSB1C3.

Protocols Used: Ligations Overnight Cultures

Results:N/A

Next Steps: The pSB1C3-hHBN ligations will be transformed into MC1061 E.coli tomorrow. The overnight cultures will be subcultured tomorrow and induced with IPTG to test protein expression levels.

14/7: Transformation of pSB1C3-hHBN into MC1061 E.coli and Optmiization of hHBA and hHBB Expression

Aim of experiment: To transform pSB1C3-hHBN into MC1061 E.coli and set up expression optimization tests for hHBA and hHBB.

Protocols Used: Transformations Protein Expression Optimization

Results: Figure 6

Next Steps: If transformations are successful, overnight cultures will be set up tomorrow. Western Blotting will be continued tomorrow for hHBA and hHBB.

15/7: Overnight Cultures of MC1061 E.coli containing pSB1C3-hHBN and Western Blotting of hHBA and hHBB

Aim of experiment: To set up overnight culture of MC1061 E.coli containing pSB1C3-hHBN and finish Western Blotting of hHBA and hHBB.

Protocols Used: Overnight Cultures Western Blotting

Results: Figure 7

Next Steps: The blot shows that characterization of hHBB has been successful. Unfortunately, nothing was visible on the blot for hHBA. 3L cultures for purification of hHBB will be set up this week. Characterization of hHBA may be reattempted. The overnight cultures will be miniprepped tomorrow.

16/7: Plasmid Purification of pSB1C3-hHBN and Overnight Cultures of M15[pREP4] E.coli containing pQE80-L-hHBB

Aim of experiment: To miniprep the MC1061 E.coli overnight cultures containing pSB1C3-hHBN and set up overnight cultures of M15[pREP4] E.coli containing pQE80-L-hHBB for 3L cultures.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit) Overnight Cultures

Results: N/A

Next Steps: The miniprep with the highest concentration will be sent for sequencing. The overnight cultures will be used to set up 3L day cultures tomorrow for purrification of hHBB.

17/7: 3L Cultures of M15[pREP4] E.coli containing pQE80-L-hHBB

Aim of experiment: To set up 3L day cultures of M15[pREP4] E.coli containing pQE80-L-hHBB for purification of hHBB.

Protocols Used: Protein Purification Note: The protocol was stopped at Step 6.

Results: N/A

Next Steps: Purification will be continued on Monday.

Week Beginning 20/7/15

Summary

hHBB was successfully purified this week and cloning of hHBN into pQE80-L was completed.

20/7: Amplification of hHBN for Cloning into pQE80-L and Purification of hHBB

Aim of experiment: To amplify hHBN for cloning into pQE80-L and continuation of hHBB purification.

Protocols Used: PCR Protocols Used: Protein Purification Note: Protocol was continued from Step 6 PCR

Results: Figure 8

Next Steps: The fractions from the affinity chromatography will be consolidated and concentrated tomorrow. Size exclusion chromatography will also be performed. The hHBN PCR product will be digested and ligated into pQE80-L tomorrow.

21/7: Size Exclusion Chromatography of hHBB and Restriction Digests and Ligations of hHBN into pQE80-L

Aim of experiment: To consolidate, concentrate and perform size exclusion chromatography on the hHBB samples. To digest the hHBN PCR product with BamHI and KpnI and subsequently ligate it into pQE80-L.

Protocols Used: Protein Purification SDS Note: An SDS gel was ran of the SEC fractions which corresponded to the observed peak. Western blotting was stopped at the blocking stage. Restriction Digests Ligations

Results: Figure 9

Next Steps: Western blotting will be completed tomorrow and will confirm if the bands observed on the SDS gel are indeed hHBB. The pQE80-LhHBN ligations will be transformed into M15[pREP4] E.coli tomorrrow.

22/7: Western Blot of hHBB and Transformations of m15[pREP4] E.coli with pQE80-LhHBN Ligations

Aim of experiment: To complete the Western blot of hHBB and transform pQE80-LhHBN into m15[pREP4] E.coli.

Protocols Used: Western Blotting Transformations

Results: Figure 10

Next Steps: The blot shows hHBB has been successfully purified. If the transformations are successful, overnight cultures will be set up tomorrow.

23/7: Overnight Cultures of m15[pREP4] E.coli containing pQE80-LhHBN

Aim of experiment: To set up overnight cultures of m15[pREP4] E.coli containing pQE80-LhHBN.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The overnight cultures will be miniprepped tomorrow.

24/7: Plasmid Purification of Overnight Cultures of m15[pREP4] E.coli containing pQE80-LhHBN

Aim of experiment: To miniprep m15[pREP4] E.coli overnight cultures containing pQE80-LhHBN.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with the highest concentration will be sent for sequencing.

Week Beginning 27/7/15

Summary

hHBN was successfully cloned into pQE80-L and cloning started for insertion into pT25.

27/7: Restriction Digests and Ligations of hHBN into pQE80-L

Aim of experiment: To digest hHBN again with BamHI and KpnI and set up ligations with pQE80-L.

Protocols Used: Resriction Digests Ligations

Results: N/A

Next Steps: Ligations will be transformed into M15[pREP4] E.coli tomorrow.

28/7: Transformation of M15[pREP4] E.coli with pQE80-L-hHBN

Aim of experiment: To transform M15[pREP4] E.coli with pQE80-L-hHBN.

Protocols Used: Transformations

Results: N/A

Next Steps: If transformations are successful, overnight cultures will be set up tomorrow.

29/7: Overnight Cultures of M15[pREP4] E.coli containing pQE80-L-hHBN

Aim of experiment: To set up overnight cultures of M15[pREP4] E.coli containing pQE80-L-hHBN

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The overnight cultures will be miniprepped tomorrow.

30/7: Plasmid Purification of M15[pREP4] E.coli Overnight Cultures containing pQE80-L-hHBN

Aim of experiment: To miniprep the M15[pREP4] E.coli Overnight Cultures containing pQE80-L-hHBN.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps:The miniprep with the highest concentration will be sent for sequencing.

31/7: Amplification of hHBN for Cloning into pT25

Aim of experiment: To amplfy hHBN for cloning into the two hybrid vector pT25.

Protocols Used: PCR

Results: Figure 11

Next Steps:The PCR product will be digested and ligated into pT25 on Monday.

Week Beginning 3/8/15

Summary

Cloning of hHBN into pT25 was completed this week but subsequent sequencing showed that it had not inserted successfully.

3/8: Restriction Digests and Ligations of hHBN for Cloning into pT25

Aim of experiment: To digest the hHBN PCR product with BamHI and KpnI and set up ligations with the pT25 vector.

Protocols Used: Restriction Digests Ligations

Results: N/A

Next Steps:The pT25-hHBN ligations will be transformed into MG1655 E.coli tomorrow.

4/8: Transformations of pT25-hHBN into MG1655 E.coli

Aim of experiment: To set up transformations of pT25-hHBN into MG1655 E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: If transformations are successful, overnight cultures will be set up tomorrow.

5/8: Overnight Cultures of MG1655 E.coli containing pT25-hHBN

Aim of experiment: To set up overnight cultures of MG1655 E.coli containing pT25-hHBN.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The overnight cultures will be miniprepped tomorrow.

6/8: Plasmid Purification of MG1655 E.coli Overnight Cultures containing pT25-hHBN

Aim of experiment: To miniprep the MG1655 E.coli overnight cultures containing pT25-hHBN.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with the highest concentration will be sent for sequencing.

Week Beginning 10/8/15

Summary

Overexpression assays of hHBN were set up this week and purification of the protein was begun.

10/8: Overnight Cultures for hHBN Overexpression Assays

Aim of experiment: To set up overnight cultures of M15[pREP4] E.coli containing pQE80-L-hHBN.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps:The overnight cultures will be used to set up day cultures that will be induced with different concentrations of IPTG to assay hHBN expression levels.

11/8: Overexpression Assay of pQE80-L-hHBN

Aim of experiment: To assay hHBN expression levels when induced with a range of IPTG concentrations.

Protocols Used: Overexpression Assay Note: Day cultures were grown to an OD₆₀₀ of 0.6 and then induced with 0.25 mM, 0.5 mM, 0.75 mM and 1 mM IPTG and allowed to grow for 6 hours.

Results: SDS gel

Next Steps: The gel indicates that hHBN is not being overexpressed in any of the cultures. Further optimization experiments will be set up to try and overexpress hHBN.

12/8: Overnight Cultures of M15[pREP4] E.coli containing pQE80-L-hHBN

Aim of experiment: To set up overnight cultures of M15[pREP4] E.coli containing pQE80-L-hHBN which will be used to set up a 3 L day culture tomorrow.

Protocols Used: Overnight Cultures Note: A 150 ml overnight culture was set up.

Results: N/A

Next Steps: The overnight cultures will be used to set up 3 L day cultures tomorrow.

13/8: 3L Day Cultures of M15[pREP4] E.coli containing pQE80-L-hHBN

Aim of experiment: To set up 3L day cultures of M15[pREP4] E.coli containing pQE80-L-hHBN using the overnight culture set up yesterday.

Protocols Used: Protein Purification Note: The cultures were grown to OD₆₀₀=0.9, then induced with 1 mM IPTG and allowed to grow for a further 8 hours. A 1 ml sample was taken before the cultures were spun down.

Results: N/A

Next Steps: Protein purification will be continued next week.

Week Beginning 17/8/15

Summary

Purification of haptoglobin was completed this week and confirmed via Western Blotting.

17/8: hHBN Purification

Aim of experiment: To continue purification of hHBN by performing affinity chromatography.

Protocols Used: Protein Purification Note: The protocol was stopped at Step 13.

Results: N/A

Next Steps:The samples corresponding to the peak observed in the chromatogram will be combined and purified using SEC tomorrow.

18/8: hHBN Purification

Aim of experiment: To continue purification of hHBN by performing size exclusion chromatography.

Protocols Used: Protein Purification

Results:

Next Steps:The gel shows bands corresponding to 45kDa, the expected size of hHBN. This will be checked via Western Blotting to ensure it is the correct protein.

19/8: Western Blotting of Purified hHBN

Aim of experiment: To perform Western Blotting in order to verify the protein that was purified is indeed hHBN.

Protocols Used: Western Blotting Note: The membrane was left blocking in milk overnight.

Results: N/A

Next Steps: The blot will be completed tomorrow.

20/8: Western Blotting of Purified hHBN

Aim of experiment: To complete Western Blotting of hHBN.

Protocols Used: Western Blotting The protocol was continued from Step 12.

Results: Figure 14

Next Steps: The blot indicates that hHBN has been successfully purified.

Week Beginning 24/8/2015

Summary

Cloning of hHBA into pT25 was carried out this week.

24/8: Amplification of hHBA for Cloning into pT25

Aim of experiment: To amplify hHBA for subsequent cloning into the two hybrid vector pT25.

Protocols Used: PCR

Results: N/A

Next Steps: The gel extracted PCR product will be digested and ligated into pT25 tomorrow.

25/8: Restriction Digests of hHBA and Ligations into pT25

Aim of experiment: To digest the hHBA with BamHI and KpnI and set up ligations with pT25.

Protocols Used: Restriction Digests Ligations

Results: N/A

Next Steps:The ligations will be left overnight and transformed into MG1065∆cya E.coli tomorrow.

26/8: Transformations of pT25-hHBA into MG1065∆cya E.coli

Aim of experiment: To transform the pT25-hHBA ligations into MG1065∆cya E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: If transformations are successful, colonies will be pick for setting up overnight cultures tomorrow.

27/8: Overnight Cultures of MG1065∆cya E.coli containing pT25-hHBA

Aim of experiment: To set up overnight cultures of MG1065∆cya E.coli containing pT25-hHBA.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The overnight cultures will be miniprepped tomorrow.

28/8: Plasmid Purification of MG1065∆cya E.coli containing pT25-hHBA Overnight Cultures

Aim of experiment: To miniprep the MG1065∆cya E.coli overnight cultures containing pT25-hHBA.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with highest concentration was sent for sequencing to confirm if the insert has been cloned in successfully.

Week Beginning 7/9/2015

Summary

Cloning of hHBN into pUT18 was carried out this week.

24/8: Amplification of hHBN for Cloning into pUT18

Aim of experiment: To amplify hHBN for subsequent cloning into the two hybrid vector pUT18.

Protocols Used: PCR

Results: N/A

Next Steps: The gel extracted PCR product will be digested and ligated into pT25 tomorrow.

25/8: Restriction Digests of hHBN and Ligations into pUT18

Aim of experiment: To digest the hHBN with BamHI and KpnI and set up ligations with pUT18.

Protocols Used: Restriction Digests Ligations

Results: N/A

Next Steps:The ligations will be left overnight and transformed into MG1065∆cya E.coli tomorrow.

26/8: Transformations of pUT18-hHBN into MG1065∆cya E.coli

Aim of experiment: To transform the pUT18-hHBN ligations into MG1065∆cya E.coli.

Protocols Used: Transformations

Results: N/A

Next Steps: If transformations are successful, colonies will be pick for setting up overnight cultures tomorrow.

27/8: Overnight Cultures of MG1065∆cya E.coli containing pUT18-hHBN

Aim of experiment: To set up overnight cultures of MG1065∆cya E.coli containing pUT18-hHBN.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: The overnight cultures will be miniprepped tomorrow.

28/8: Plasmid Purification of MG1065∆cya E.coli containing pUT18-hHBN Overnight Cultures

Aim of experiment: To miniprep the MG1065∆cya E.coli overnight cultures containing pUT18-hHBN.

Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit)

Results: N/A

Next Steps: The miniprep with highest concentration was sent for sequencing to confirm if the insert has been cloned in successfully.

@@ Line 12: / Line 12: @@
        </header>
-<body>
+<!-- button navigation -->
+<a class="anchor" id="selection"></a>
+    <section id="about" class="row1">
+      <div class="row">
+        <div class="col-lg-4">
+          <a href="#section1" class="scroll"><span class="glyphicon glyphicon-briefcase"></span></a>
+          <h3>BioSpray</h3>
+          <p class="about-content">Our forensic toolkit aims to use synthetic biology approaches to improve on the current
+methods used by crime scene investigators.</p>
+        </div>
+        <div class="col-lg-4">
+          <a href="#section2"><span class="glyphicon glyphicon-search" type="button"></span></a>
+          <h3>Chromium Detector</h3>
+          <p class="about-content">Lorem ipsum dolor sit amet, consectetur adipiscing elit. Curabitur a tincidunt elit. Aliquam porta nibh at enim luctus, auctor consequat dolor vehicula.</p>
+        </div>
+        <div class="col-lg-4">
+          <a href="#section3"><span class="glyphicon glyphicon-eye-open"></span></a>
+          <h3>Fingerprint Aging</h3>
+          <p class="about-content">Lorem ipsum dolor sit amet, consectetur adipiscing elit. Curabitur a tincidunt elit. Aliquam porta nibh at enim luctus, auctor consequat dolor vehicula.</p>
+        </div>
+      </div>
+    </section>
+<div class="ribbon">
+<div class="rows">
+<!-- SALIVA-->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <span id="biospray-title">Saliva</span>
+          <span type="button" class="plusminus-toggleable box-btn glyphicon glyphicon glyphicon-plus-sign" data-toggle="collapse" data-target="#sacollapsible"></span>
+           <div id="sacollapsible" class="collapse biospray-content">
+<!-- WEEK 1 -->
+  <div class="row">
+     <div class="box">
+        <div class="labtitle">Week Beginning 1/6/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew1"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content"><i>Lactoferrin binding protein A (LbpA)</i> was successfully cloned into the vector pSB1C3.</p>
+         </div>
+         <div id="sacollapsiblew1" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">1/6: Amplification of <i>LbpA</i> for Cloning into pSB1C3</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew1-1"></span>
+                <div id="sacollapsiblew1-1" class="collapse box-content">
+                  <p><b>Aim of Experiment:</b> Amplify <i>LbpA</i> from MC58 <i>N.meningitidis</i> template chromosome.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR</a> </p>
+         <p><b>Results: </b><a data-toggle="modal" data-target="#lbpa-figure1-modal" class="journal-protocol"> Figure 1 </a></p>
+                  <p><b>Next Steps:</b> Since the <i>LbpA</i> gene was amplified successfully, it was ready to be digested for subsequent cloning into pSB1C3.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">2/6: Restriction Digests and Ligations with pSB1C3</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew1-2"></span>
+                <div id="sacollapsiblew1-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest <i>LbpA</i> with BamHI and EcoRI and then ligate it into pSB1C3.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol">Restriction Digests</a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a></p>
+         <p><b>Results:</b> N/A
+                  <p><b>Next Steps:</b> Ligations of pSB1C3-<i>LbpA</i> were left overnight until the next day when they would be transformed into the JM110 <i>E.coli</i>.
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">3/6: Transformations of pSB1C3-<i>LpbA</i> into JM110 <i>E.coli</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew1-3"></span>
+                <div id="sacollapsiblew1-3" class="collapse box-content">
+                 <p><b>Aim of experiment:</b> To transform the pSB1C3-<i>LbpA</i> ligations into JM110 <i>E.coli</i>.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a></p>
+                  <p><b>Results:</b> N/A
+                  <p><b>Next Steps:</b> Transformations will be checked tomorrow and if successful, overnight cultures will be set up.
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">4/6: Overnight Cultures of JM110 <i>E.coli</i> containing pSB1C3-<i>LbpA</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew1-4"></span>
+                <div id="sacollapsiblew1-4" class="collapse box-content">
+                 <p><b>Aim of experiment:</b> To set up overnight cultures of JM110 <i>E.coli</i> containing pSB1C3-<i>LbpA</i>.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures
+          </a></p>
+                  <p><b>Results:</b> N/A
+                  <p><b>Next Steps:</b> A miniprep will be performed on the overnight cultures and a pre-sequence digest test performed tomorrow.
+                </div>
+        </div>
+        </div>
+      </div>
+            <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">5/6: Plasmid Purification and Pre-Sequencing Digest Check</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew1-5"></span>
+                <div id="sacollapsiblew1-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the JM110 <i>E.coli</i> overnight cultures containing pSB1C3-<i>LbpA</i>.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit) </a>
+          </a></p>
+                  <p><b>Results:</b> N/A
+                  <p><b>Next Steps:</b> The miniprep with the highest concentration will be sent for sequencing.                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+  </div>
+<!-- WEEK 2 -->
+  <div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 8/6/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew2"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Sequencing confirmed that <i>LbpA</i> had been successfully cloned into pSB1C3 therefore this week cloning was attempted to put <i>LbpA</i> into the high expression vector pQE80-L.</p>
+         </div>
+         <div id="sacollapsiblew2" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">8/6: Amplification of <i>LbpA</i> for Cloning into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew2-1"></span>
+                <div id="sacollapsiblew2-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplify <i>LbpA</i> for subsequent cloninng into the pQE80-L vector.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR </a></p>
+                  <p><b>Results:</b><a data-toggle="modal" data-target="#lbpa-figure2-modal" class="journal-protocol"> Figure 2 </a></p>
+<p><b> Next Steps:</b> The PCR product will be digested and ligated into pQE80-L tomorrrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">9/6: Restriction Digests and Ligations of <i>LbpA</i>into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew2-2"></span>
+                <div id="sacollapsiblew2-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To restrict and ligate the <i>LbpA</i> gene into the pQE80-L plasmid.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol">Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations </a> </p>
+              <p><b> Results:</b>: N/A </p>
+<p><b> Next Steps:</b> Transformations of pQE80-L-<i>LbpA</i> into JM110 <i>E.coli</i> will be carried out tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">10/6: Transformations of pQE80-L-<i>LpbA</i> into JM110 <i>E.coli</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew2-3"></span>
+                <div id="sacollapsiblew2-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform  pQE80-L-<i>LpbA</i> into JM110 <i>E.coli</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a></p>
+              <p><b>Results:</b> N/A </p>
+<p><b> Next Steps:</b> If the transformations are successful, colonies will be picked to set up overnight cultures. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 3 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 15/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew3"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">This week, cloning of <i>LbpA</i> into pQE80-L was continued due to last week's failed attempts.</p>
+         </div>
+         <div id="sacollapsiblew3" class="collapse week-content">
+     <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">15/6: PCR of <i>LbpA</i> for Cloning into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew3-1"></span>
+                <div id="sacollapsiblew3-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To re-amplify the <i>LbpA</i> gene for cloning into the pQE80-L vector.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR</a></p>
+                  <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure3-modal" class="journal-protocol">Figure 3</a></b> </p>
+<p><b>Next Steps:</b> The gel shows that <i>LbpA</i> has been successfully amplified so it will be digested and ligated into the pQE80-L vector tomorrow.  </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 4 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 22/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew4"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content"><i>LbpA</i> was successfully cloned into the high expression vector pQE80-L.</p>
+         </div>
+         <div id="sacollapsiblew4" class="collapse week-content">
+     <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">23/6: Restriction Digest and Ligation of <i>LbpA</i> into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew4-1"></span>
+                <div id="sacollapsiblew4-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest the amplified <i>LbpA</i> gene with BamHI and KpnI and subsequently ligate it into the pQE80-L vector.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a></p>
+                  <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The ligations will be transformed into M15pREP4 <i>E.coli</i>. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">24/6: Transformations of pQE80-L-<i>LbpA</i> into M15pREP4 <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew4-2"></span>
+                <div id="sacollapsiblew4-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform pQE80-L-<i>LbpA</i> into M15pREP4 <i>E.coli</i>. </p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations</a></p>
+                  <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Overnight cultures will be set up tomorrow if the transformations are successful. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">25/6: Overnight Cultures of M15pREP4  <i>E.coli</i> containing pQE80-L-<i>LbpA</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew4-3"></span>
+                <div id="sacollapsiblew4-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of M15pREP4 <i>E.coli</i> containing pQE80-L-<i>LbpA</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures </a></p>
+                  <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Overnight cultures will be miniprepped and a pre-sequence restriction digest will be performed to check for the presence of the <i>LbpA</i> gene. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">26/6: Plasmid Purification of the M15pREP4 <i>E.coli</i> Overnight Cultures containing pQE80-L-<i>LbpA</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew4-4"></span>
+                <div id="sacollapsiblew4-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the overnight cultures from yesterday and perform a pre-sequencing restriction digest using BamHI and KpnI to check for presence of <i>LbpA</i> gene.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol">Plasmid Purification (QIAprep® Spin Miniprep Kit)</a></p>
+                  <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> A sample of the miniprep was sent for sequencing to confirm the presence of <i>LbpA</i> in the pQE80-L vector. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 5 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 29/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew5"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Protein expression optimization experiments were performed on LbpA to assay protein production levels. However, cell growth seems to cease when <i>LbpA</i> expression is induced.</p>
+         </div>
+         <div id="sacollapsiblew5" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">1/7: M15pREP4 <i>E.coli</i> Overnight Cultures containing pQE80-L-<i>LbpA</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew5-1"></span>
+                <div id="sacollapsiblew5-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures in preparation for protein expression experiments tomorrow.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a></p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The overnight cultures will be used to set up day cultures tomorrow which will be used to test expression of <i>LbpA</i>. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">2/7: Optimization of <i>LbpA</i> Expression</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew5-2"></span>
+                <div id="sacollapsiblew5-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up day cultures and induce expression of <i>LbpA</i> using different concentrations of IPTG and check levels of protein production via SDS-PAGE.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Optimization </a> </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> The samples were stored in the -20 <sup>o</sup>C freezer overnight and will be ran on a gel tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">3/7: Continuation of Optimization of <i>LbpA</i> Expression</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew5-3"></span>
+                <div id="sacollapsiblew5-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To test the samples obtained yesterday on an SDS gel.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Optimization </a> </p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure4-modal" class="journal-protocol"> Figure 4 </a> </p>
+<p><b>Next Steps:</b> Further optimization experiments will be set up next week using different concentrations of IPTG. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 6 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 6/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew6"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Further protein expression optimization experiments were set up, this time using a range of IPTG concentrations to induce expression of <i>LbpA</i>.</p>
+         </div>
+         <div id="sacollapsiblew6" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">7/7: Further Optimization of <i>LbpA</i> Expression</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew6-1"></span>
+                <div id="sacollapsiblew6-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up further tests to optimize expression of LbpA using different concentrations of IPTG.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Optimization </a> Note: An SDS gel was not ran, instead the OD<sub>600</sub> readings were recorded for the uninduced and induced cultures at 20 minute intervals for 2 hours, 1 and half hours after the cultures were induced. </p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure5-modal" class="journal-protocol"> Figure 5 </a> </p>
+<p><b>Next Steps:</b> A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after <i>LbpA</i> expression has been induced. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 7 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 13/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew7"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Protein expression optimization experiments were continued this week. A growth curve assay was carried out to assess the effect of <i>LbpA</i> expression over a longer period of time.  </p>
+         </div>
+         <div id="sacollapsiblew7" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">14/7: Plate Reader Growth Curve Assay</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew7-1"></span>
+                <div id="sacollapsiblew7-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To induce expression of <i>LbpA</i> using different concentration of IPTG and allow cells to grow for 16 hours in order to monitor cell growth.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#platereader-modal" class="journal-protocol"> Growth Curve Assay </a></p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure6-modal" class="journal-protocol"> Figure 6 </a> </p>
+<p><b>Next Steps:</b> A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after <i>LbpA</i> expression has been induced. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">17/7: Sample Preparation for Western Blot</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew7-2"></span>
+                <div id="sacollapsiblew7-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To check if the cells in the induced cultures ~7 hours after being induced are producing LbpA. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blot Sample Preparation </a> Note: The day cultures were grown for 7 hours after they were induced and then the samples were frozen, to be blotted on Monday. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The samples will be ran on an SDS gel on Monday and blotted. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 8 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 20/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew8"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Samples from the growth curve assay were blotted to see if LbpA is being produced at the later stages of the growth curve. A further growth curve assay was also performed which monitored cell growth until a certain OD<sub>600</sub> was reached, at which point the cultures were induced.</p>
+         </div>
+         <div id="sacollapsiblew8" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">20/7: Western Blot LpbA Culture Samples</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew8-1"></span>
+                <div id="sacollapsiblew8-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To western blot the LbpA culture samples from Friday.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blot </a></p>
+              <p><b>Results:</b> Nothing was visible on the blot. </p>
+<p><b>Next Steps:</b> Another plate reader experiment will be set up on Thursday where the cultures will be monitored for three hours before induction with IPTG and then for a further 20 hours afterwards. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">23/7: Growth Curve Assay</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew8-2"></span>
+                <div id="sacollapsiblew8-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To check if the cells begin to die after induction with IPTG after having already grown to an OD<sub>600</sub> of 0.5. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#platereader2-modal" class="journal-protocol"> Growth Curve Assay </a> </p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure7-modal" class="journal-protocol"> Figure 7 </a> </p>
+<p><b>Next Steps:</b> Samples of these cultures will be taken 6 hours after induction to check for the presence of LbpA. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 9 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 27/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew9"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">The presence of LbpA was confirmed in the culture 6 hours after induction with IPTG so protein purification was started this week.</p>
+         </div>
+         <div id="sacollapsiblew9" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">27/7: Overnight Cultures of M15pREP4 <i>E.coli</i> containing pQE80-L-<i>LbpA</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew9-1"></span>
+                <div id="sacollapsiblew9-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up a 150 ml overnight culture for sub-culturing tomorrow.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> Note: 150ml of LB was used and 150µl of each antibiotic. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>150ml of fresh LB will be inoculated tomorrow with 7.5 ml of the overnight cultures. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">28/7: <i>LbpA</i> Expression Assay</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew9-2"></span>
+                <div id="sacollapsiblew9-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up day cultures using the overnight cultures from yesterday and test for expression of LbpA 6 hours after induction.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Test </a> Note: 150 ml of fresh LB was inoculated with 7.5 ml of the overnight cultures. Only an uninduced control and an 1 mM IPTG induced culture was set up. The samples were not blotted, they were only ran on an SDS gel. </p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure8-modal" class="journal-protocol"> Figure 8 </a>   </p>
+<p><b>Next Steps:</b> A 3 L culture will be set up on Thursday under the same conditions as the overnight cultures in order to try and purify LbpA.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">29/7: Overnight Cultures of M15pREP4 <i>E.coli</i> containing pQE80-L-<i>LbpA</i> for 3L Culture</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew9-3"></span>
+                <div id="sacollapsiblew9-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures for the 3L day cultures that will be grown tomorrow.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> Note: One 150 ml overnight culture was set up using 150 µl of each antibiotic. </p>
+              <p><b>Results:</b> N/A   </p>
+<p><b>Next Steps:</b> The 3 L culture will be set up tomorrow allowed to grow to OD<sub>600</sub> of 0.6-1 and then induced with 1 mM IPTG and grown for 6 hours.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">30/7: 3 L Culture of M15pREP4 <i>E.coli</i> containing pQE80-L-<i>LbpA</i> for Purification of LbpA</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew9-4"></span>
+                <div id="sacollapsiblew9-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up 3 L day cultures for purifying LbpA.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: The procedure was stopped at Step 6 and the pellets were frozen at -20 <sup>o</sup>C. </p>
+              <p><b>Results:</b> N/A   </p>
+<p><b>Next Steps:</b> Protein purification will be continued on Monday.</p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 10 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 3/8/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew10"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">The first round of LbpA purification failed at the affinity chromatography stage so another 6 L culture was grown this week to attempt purification again.</p>
+         </div>
+         <div id="sacollapsiblew10" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">3/8: Continuation of LbpA Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew10-1"></span>
+                <div id="sacollapsiblew10-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue with purification of LbpA.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: The protocol was continued from Step 7</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>Size exclusion chromatography was not carried out since an SDS gel showed LbpA was not present after performing affinity chromatography. Another 6L culture will be set up this week.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">5/8: Overnight Cultures of M15pREP4 <i>E.coli</i> containing pQE80-L-<i>LbpA</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew10-2"></span>
+                <div id="sacollapsiblew10-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures for the 6 L culture that will be set up tomorrow.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a>Note: One 150 ml overnight culture was set up using 150 µl of each antibiotic.</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>The overnight cultures will be used to set up the 6 L day culture tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">6/8: 6 L Day Cultures of M15pREP4 <i>E.coli</i> containing pQE80-L-<i>LbpA</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew10-3"></span>
+                <div id="sacollapsiblew10-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up 6 L day cultures for purifying LbpA and begin the purification process.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: The protocol was stopped at Step 6.</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>Purification of LbpA will be continued on Monday. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 11 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 10/8/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew11"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Purification of LbpA was continued but unfortunately was unsuccessful. If time permits, another round of purification may be reattempted. </p>
+         </div>
+         <div id="sacollapsiblew11" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">10/8: Continuation of LbpA Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew11-1"></span>
+                <div id="sacollapsiblew11-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue with purification of LbpA.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: The protocol was continued from Step 7</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Purification of LbpA will be continued tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">11/8: Continuation of LbpA Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sacollapsiblew11-2"></span>
+                <div id="sacollapsiblew11-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To purify LbpA using affinity chromatography.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: The protocol was stopped at Step 13. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>Since no peaks were observed, purification of LbpA was abadoned at this stage. Purification may be retried. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+         </div>
+       </div>
+   </div>
+</div>
+<!-- SEMEN-->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <span id="biospray-title">Semen</span>
+          <span type="button" class="plusminus-toggleable box-btn glyphicon glyphicon glyphicon-plus-sign" data-toggle="collapse" data-target="#secollapsible"></span>
+           <div id="secollapsible" class="collapse biospray-content">
+<!-- WEEK 1 -->
+  <div class="row">
+     <div class="box">
+        <div class="labtitle">Week Beginning 25/5/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew1"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i> PotD</i> into the biobrick vector, pSB1C3, was started this week.</p>
+         </div>
+         <div id="secollapsiblew1" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">28/5: PCR of <i>PotD</i> from MG1655 <i>E.coli</i> gDNA</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew1-1"></span>
+                <div id="secollapsiblew1-1" class="collapse box-content">
+                  <p><b>Aim of Experiment:</b> To amplify <i>PotD</i> from MG1655 <i>E.coli</i> gDNA.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR</a></p>
+         <p><b>Results: </b>N/A</p>
+                  <p><b>Next Steps:</b> The PCR product will be gel extracted, digested and ligated into the biobrick vector pSB1C3 tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">29/5: Gel Extraction, Restriction Digest and Ligation of <i>SBP</i> into pSB1C3</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew1-2"></span>
+                <div id="secollapsiblew1-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To gel extract the <i>PotD</i> PCR product and digest it with PstI and EcoRI. A ligation will then be set up with pSB1C3.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol">Restriction Digest</a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol">Ligation</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> Ligations will be transformed into JM110 <i>E.coli</i> on Monday.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+  </div>
+<!-- WEEK 2 -->
+  <div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 1/6/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew2"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>PotD</i> into the biobrick vector was completed this week. </p>
+         </div>
+         <div id="secollapsiblew2" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">1/6: Transformation of pSB1C3-<i>PotD</i> into JM110 <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew2-1"></span>
+                <div id="secollapsiblew2-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform pSB1C3-<i>PotD</i> into JM110 <i>E.coli</i>.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a></p>
+                  <p><b>Results:</b>N/A</p>
+<p><b> Next Steps:</b> If the transformations are successful, overnight cultures will be set up tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">2/6: Overnight Cultures of pSB1C3-<i>PotD</i> in JM110 <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew2-2"></span>
+                <div id="secollapsiblew2-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of pSB1C3-<i>PotD</i> in JM110 <i>E.coli</i></p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures </a></p>
+              <p><b> Results:</b>: N/A </p>
+<p><b> Next Steps:</b> The overnight cultures will be miniprepped tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">3/6: Plasmid Purification of pSB1C3-<i>PotD</i> from Overnight Cultures</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew2-3"></span>
+                <div id="secollapsiblew2-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the overnight cultures from yesterday.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit) </a></p>
+              <p><b>Results:</b> N/A </p>
+<p><b> Next Steps:</b> The miniprep with the highest concentration will be sent for sequencing to confirm <i>PotD</i> has been inserted into pSB1C3 successfully.</p>
+                 </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 3 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 15/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">This week, cloning of <i>PotD</i> into pQE80-L was started. The gene for <i>Spermine Binding Protein (SBP) </i> also arrived in a plasmid so it was transformed into MC1061 <i>E.coli</i> and plasmid purified.</p>
+         </div>
+         <div id="secollapsiblew3" class="collapse week-content">
+     <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">15/6: PCR of <i>PotD</i> for Cloning into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3-1"></span>
+                <div id="secollapsiblew3-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplify the <i>PotD</i> gene for cloning into the pQE80-L vector.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR</a></p>
+                  <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure3-modal" class="journal-protocol">Figure 3</a></b> </p>
+<p><b>Next Steps:</b> The gel shows that <i>PotD</i> has been successfully amplified so it will be digested and ligated into the pQE80-L vector tomorrow.  </p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">16/6: Gel Extraction of <i>PotD</i> PCR Product</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3-2"></span>
+                <div id="secollapsiblew3-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To gel extract the <i>PotD</i> PCR product.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR</a></p>
+                  <p><b>Results:</b> <a data-toggle="modal" data-target="#lbpa-figure3-modal" class="journal-protocol">Figure 3</a></b> </p>
+<p><b>Next Steps:</b> The gel shows that <i>PotD</i> has been successfully amplified so it will be digested and ligated into the pQE80-L vector tomorrow.  </p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">17/6: Restriction Digest and Ligation of <i>PotD</i> into pQE80-L. Transformation of IDT Plasmid containing <i>SBP</i> into MC1061 <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3-3"></span>
+                <div id="secollapsiblew3-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest the <i>PotD</i> PCR product with KpnI and BamHI and then ligate it into pQE80-L. To transform MC1061 <i>E.coli</i> with the IDT plasmid containing <i>SBP</i>.</p>
+                  <p><b>Protocols Used: </b></p><i>PotD</i>: <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests</a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a> <i>SBP</i>:<a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol">Transformations</a> </p>
+                  <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> The pQE80-L-<i>PotD</i> ligations will be transformed into MC1061 <i>E.coli</i>. Overnight cultures of the transformations will be set up tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">18/6: Transformations of <i>PotD</i> + pQE80-L into MC1061 <i>E.coli</i>. Overnight Cultures of MC1061 <i>E.coli</i> with SBP + IDT Plasmid</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3-4"></span>
+                <div id="secollapsiblew3-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform the pQE80-L-<i>PotD</i> ligations into MC1061 <i>E.coli</i>. To set up overnight cultures of the transformations of MC1061 <i>E.coli</i> with <i>SBP</i> + IDT Plasmid that were set up yesterday.</p>
+                  <p><b>Protocols Used: </b><i>PotD</i>:<a data-toggle="modal" data-target="#transformations-modal" class="journal-protocol"> Transformations</a> <i>SBP</i>: <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures</a></p>
+                  <p><b>Results:</b>N/A </p>
+<p><b>Next Steps:</b> If transformations are successful, overnight cultures will be set up tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">19/6: Repeat of Ligation of <i>PotD</i> into pQE80-L. Plasmid Purification of the IDT Plasmid Containing <i>SBP</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3-5"></span>
+                <div id="secollapsiblew3-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To repeat ligations of <i>PotD</i> into pQE80-L since transformations did not work. To miniprep the overnight cultures that were set up yesterday.</p>
+                  <p><b>Protocols Used: </b><i>PotD</i>: <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a> <i>SBP</i>: <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a></p>
+                  <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Ligations will be left over the weekend and transformed on Monday. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 4 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 22/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew4"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>PotD</i> into the high expression vector pQE80-L was completed. Cloning of <i>SBP</i> into pSB1C3 and pQE80-L was started this week.</p>
+         </div>
+         <div id="secollapsiblew4" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">22/6: Transformation of pQE80-L-<i>PotD</i> Ligations. PCR and Gel Extraction of <i>SBP</i> for Cloning into pQE80-L and Restriction Digests and Ligation of <i>SBP</i> for Cloning into pSB1C3</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew4-1"></span>
+                <div id="secollapsiblew4-1" class="collapse box-content">
+                  <p><b>Aim of Experiment:</b> To transform pQE80-L-<i>PotD</i> into  MC1061<i>E.coli</i>. To set up a PCR of <i>SBP</i> using the purified plasmid from Friday and to gel extract the PCR product for cloning into pQE80-L. To digest <i>SBP</i> out of the same plasmid using EcoRI and PstI and subsequently ligate it into pSB1C3.</p>
+                  <p><b>Protocols Used:</b> <i>PotD</i>: <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations</a> <i>SBP</i>: <a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR</a> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests</a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a></p>
+         <p><b>Results: </b><i>SBP</i> :<a data-toggle="modal" data-target="#sbp-figure1-modal" class="journal-protocol"> Figure 1 </a></p>
+                  <p><b>Next Steps:</b> Overnight cultures will be set up tomorrow for the <i>PotD</i> transformations. The <i>SBP</i> PCR product will be digested and ligated into pQE80-L tomorrow. The pSB1C3-<i>SBP</i> ligations will be transformed into MC1061 <i>E.coli</i>. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">23/6: Overnight Cultures of MC1061 <i>E.coli</i> containing pSB1C3-<i>PotD</i>. Transformations of MC1061 <i>E.coli</i> with pSB1C3 + <i>SBP</i>. Restriction Digests and Ligations of <i>SBP</i> into pQE80-L and Transformations into MC1061 <i>E.coli</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew4-2"></span>
+                <div id="secollapsiblew4-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of MC1061 <i>E.coli</i> containing pSB1C3 + <i>PotD</i>. To transform MC1061 <i>E.coli</i> with pSB1C3 + <i>SBP</i>. To digest the <i>SBP</i> PCR product with BamHI and Kpn I, ligate it into pQE80-L and transform these ligations into MC1061 <i>E.coli</i>.</p>
+                  <p><b>Protocols Used:</b> <i>PotD</i>: <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> <i>SBP</i>: <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a><a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations </a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The MC1061 <i>E.coli</i> overnight cultures containing pQE80-L + <i> PotD</i> will be miniiprepped tomorrow. Overnight cultures will be set up of the MC1061 <i>E.coli</i> transformations with pSB1C3 + <i>SBP</i> and pQE80-L + <i>SBP</i>.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">24/6: Plasmid Purification of the Overnight Cultures of MC1061 <i>E.coli</i> containing pSB1C3-<i>PotD</i>. Overnight Cultures of the Transformations of MC1061 <i>E.coli</i> with pSB1C3 + <i>SBP</i> and pQE80-L + <i>SBP</i>.</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew4-3"></span>
+                <div id="secollapsiblew4-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the overnight cultures of MC1061 <i>E.coli</i> containing pSB1C3-<i>PotD</i>. To set up overnight cultures of the transformations of MC1061 <i>E.coli</i> with pSB1C3-<i>SBP</i> and pQE80-L-<i>SBP</i></p>
+                  <p><b>Protocols Used:</b> <i>PotD</i>: <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> <i>SBP</i>: <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The miniprep with the highest concentration for pQE80-L + <i>PotD</i> was sent for sequencing. The <i>SBP</i> overnight cultures will be miniprepped tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">25/6: Plasmid Purification of the Overnight Cultures of MC1061 <i>E.coli</i> containing pSB1C3-<i>SBP</i> and pQE80-L-<i>SBP</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew4-4"></span>
+                <div id="secollapsiblew4-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the overnight cultures of MC1061 <i>E.coli</i> containing pSB1C3-<i>SBP</i> and pQE80-L + <i>SBP</i>.</p>
+                  <p><b>Protocols Used:</b> <i>SBP</i>: <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit) </a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The miniprep with the highest concentration for both pSB1C3-<i>SBP</i> and pQE80-L-<i>SBP</i> were sent for sequencing. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 5 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 29/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew5"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Sequencing showed <i> SBP</i> failed to insert successfully into pSB1C3 so cloning was repeated from the ligation stage this week. However, SBP and PotD were successfully inserted into the high expression vector pQE80-L, so overexpression assays were started this week for both.</p>
+         </div>
+         <div id="secollapsiblew5" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">29/6: Ligations of <i>SBP</i> into pSB1C3 and Transformations into MC1061 <i>E.coli</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew5-1"></span>
+                <div id="secollapsiblew5-1" class="collapse box-content">
+                  <p><b>Aim of Experiment:</b> To ligate <i>SBP</i> into pSB1C3 and transform the ligations into MC1061 <i>E.coli</i>. </p>
+                  <p><b>Protocols Used:</b> <i>SBP</i>: <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations</a></p>
+         <p><b>Results: </b><i>SBP</i> :<a data-toggle="modal" data-target="#sbp-figure1-modal" class="journal-protocol"> Figure 1 </a></p>
+                  <p><b>Next Steps:</b> Overnight cultures will be set up tomorrow if transformations are successful. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">30/6: Overnight Cultures of MC1061 <i>E.coli</i> containing pSB1C3 + <i>SBP</i>, pQE80-L + <i>SBP</i> and pQE80-L + <i>PotD</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew5-2"></span>
+                <div id="secollapsiblew5-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of MC1061 <i>E.coli</i> containing pSB1C3 + <i>SBP</i>. To set up overnight cultures  of pQE80-L + <i>SBP</i> and pQE80-L + <i>PotD</i> in MC1061 <i>E.coli</i> for overexpression assays to be performed tomorrow. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The MC1061 <i>E.coli</i> overnight cultures containing pSB1C3 + <i> SBP</i> will be miniiprepped tomorrow. The MC1061 <i>E.coli</i> overnight cultures containing pQE80-L + <i> SBP</i> and pQE80-L + <i>PotD</i> will be subcultured tomorrow in order to perform overexpression assays.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">1/7: Plasmid Purification of the Overnight Cultures of MC1061 <i>E.coli</i> containing pSB1C3 + <i>SBP</i>. <i>PotD</i> and <i>SBP</i> Expression Optimization</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew5-3"></span>
+                <div id="secollapsiblew5-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the overnight cultures of MC1061 <i>E.coli</i> containing pSB1C3 + <i>SBP</i>. To start protein expression optimization for PotD and SBP.</p>
+                  <p><b>Protocols Used:</b> <i>SBP</i>: <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> <i>SBP</i> and <i>PotD</i>: <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Optimization</a> Note: Protocol was stopped at Step 8. Samples will be ran on an SDS gel tomorrow.</p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The miniprep with the highest concentration for pSB1C3 + <i>SBP</i> was sent for sequencing. The PotD and SBP samples will be ran on an SDS gel tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">2/7: Continuation of <i>PotD</i> and <i>SBP</i> Expression Optimization</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew5-4"></span>
+                <div id="secollapsiblew5-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To run the samples of PotD and SBP on an SDS gel and begin Western Blotting.</p>
+                  <p><b>Protocols Used:</b> <i>PotD</i> and <i>SBP</i>: <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Optimization</a> Note: Protocol was continued from Step 9.</p>
+         <p><b>Results:</b> <a data-toggle="modal" data-target="#sbp-figure2-modal" class="journal-protocol"> Figure 2 </a></p>
+       <p><b>Next Steps:</b> The membrane for the Western blot has been left at the blocking stage so will be continued tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">3/7: Western Blot of <i>PotD</i> and <i>SBP</i> Day Culture Samples</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew5-5"></span>
+                <div id="secollapsiblew5-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue the Western Blot of PotD and SBP.</p>
+                  <p><b>Protocols Used:</b> <i>PotD</i> and <i>SBP</i>: <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blot</a></p>
+         <p><b>Results:</b> <a data-toggle="modal" data-target="#sbp-figure3-modal" class="journal-protocol"> Figure 3 </a></p>
+       <p><b>Next Steps:</b> PotD has been successfully characterized so purification of this protein will be started next week. Further expression optimization experiments for SBP will be set up next week using different concentrations of IPTG, since its characterization was unsuccessful.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 6 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 6/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew6"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Further protein expression optimization experiments were set up, this time using a range of IPTG concentrations to induce expression of <i>SBP</i>.</p>
+         </div>
+         <div id="secollapsiblew6" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">6/7: Further Optimization of <i>SBP</i> Expression</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew6-1"></span>
+                <div id="secollapsiblew6-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up further tests to optimize expression of <i>SBP</i> using different concentrations of IPTG.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Optimization </a> Note: Cultures were induced with o.5mM, 1mM and 2mM IPTG. The Western Blot was left to block overnight. </p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#sbp-figure4-modal" class="journal-protocol"> Figure 4 </a> </p>
+<p><b>Next Steps:</b> The Western Blot will be completed tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">7/7: Western Blot of <i>SBP</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew6-2"></span>
+                <div id="secollapsiblew6-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To complete the Western Blot that was started yesterday. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blot </a> Note: Protocol was continued from Step 12.</p>
+              <p><b>Results:</b> Nothing was visible on the blot. </p>
+<p><b>Next Steps:</b> A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after <i>SBP</i> expression has been induced.  </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 7 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 13/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew7"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Protein expression optimization experiments were continued this week. A growth curve assay was carried out to assess the effect of <i>SBP</i> expression over a longer period of time. Purification of <i>PotD</i> was also started this week. </p>
+         </div>
+         <div id="secollapsiblew7" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">14/7: Plate Reader Growth Curve Assay</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew7-1"></span>
+                <div id="secollapsiblew7-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To induce expression of <i>SBP</i> using different concentration of IPTG and allow cells to grow for 16 hours in order to monitor cell growth.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#platereader-modal" class="journal-protocol"> Growth Curve Assay </a></p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#sbp-figure5-modal" class="journal-protocol"> Figure 5 </a> </p>
+<p><b>Next Steps:</b> A plate reader experiment will be set up next week to monitor the growth rate of the the cells over a longer period of time after <i>SBP</i> expression has been induced. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">15/7: Overnight Cultures for 3L Culture for PotD Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew7-2"></span>
+                <div id="secollapsiblew7-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures that will be used tomorrow to set up 3L cultures for <i>PotD</i> purification. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a></p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The 3L day culture will be set up tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">16/7: Sample Preparation for Western Blot of SBP and 3L Day Culture for PotD Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew7-3"></span>
+                <div id="secollapsiblew7-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To check if the cells in the induced cultures ~7 hours after being induced are producing SBP. To set up 3L Day Culture for <i> PotD</i> Purification </p>
+                  <p><b>Protocols Used: </b> <i>SBP</i>: <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blot Sample Preparation </a> Note: The day cultures were grown for 7 hours after they were induced and then the samples were frozen, to be blotted on Monday. <i>PotD</i>:  <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a>Note: Cultures were left to grow overnight after being induced. </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> The samples will be ran on an SDS gel on Monday and blotted. The <i>PotD</i> cultures will be harvested tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">17/7: Harvesting <i>PotD</i> Cultures</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew7-4"></span>
+                <div id="secollapsiblew7-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To harvest the day cultures grown yesterday for <i>PotD</i> purification. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: After the cultures were spun down the pellets were frozen. </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> Purification of <i>PotD</i> will continue on Monday.  </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 8 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 20/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew8"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Samples from the growth curve assay were blotted to see if SBP is being produced at the later stages of the growth curve. A further growth curve assay was also performed which monitored cell growth until a certain OD<sub>600</sub> was reached, at which point the cultures were induced. Purification of PotD was also continued this week.</p>
+         </div>
+         <div id="secollapsiblew8" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">20/7: Western Blot <i>SBP</i> Culture Samples</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew8-1"></span>
+                <div id="secollapsiblew8-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To western blot the SBP culture samples from Friday. To continue purfying PotD.</p>
+                  <p><b>Protocols Used: </b> <i>SBP</i>: <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blot </a>                           <i>PotD</i>: <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: Affinity chromatography was completed but no peaks were observable. </p>
+              <p><b>Results:</b> Nothing was visible on the blot. </p>
+<p><b>Next Steps:</b> Another plate reader experiment will be set up on Thursday where the cultures will be monitored for three hours before induction with IPTG and then for a further 20 hours afterwards. Another 150ml overnight culture was set up for <i>PotD</i> so 3L cultures could be set up tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">21/7: 3L Day Cultures for PotD Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew8-2"></span>
+                <div id="secollapsiblew8-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up 3L Day Culture for PotD Purification. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: Cultures were left to grow overnight after being induced. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Purification will be continued tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">22/7: Continuation of PotD Purification </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew8-3"></span>
+                <div id="secollapsiblew8-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue purification of PotD. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: The protocol was continued from Step 4 and stopped at Step 13.</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Purification will be continued tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">23/7: Continuation of PotD Purification and Growth Curve Assay for <i>SBP</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew8-4"></span>
+                <div id="secollapsiblew8-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue purification of PotD and to check if the cells begin to die after inducing expression of <i>SBP</i> with IPTG after having already grown to an OD<sub>600</sub> of 0.5. </p>
+                  <p><b>Protocols Used: </b> <i>PotD</i>: <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: The protocol was continued from Step 13. <i>SBP</i>: <a data-toggle="modal" data-target="#platereader2-modal" class="journal-protocol"> Growth Curve Assay </a></p>
+              <p><b>Results:</b><a data-toggle="modal" data-target="#sbp-figure6-modal" class="journal-protocol"> Figure 6 </a> </p>
+<p><b>Next Steps:</b>The samples from the size exclusion chromatography performed on PotD will be prepared tomorrow for blotting next week. For <i> SBP</i> further overexpression tests will be set up next week.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">24/7: SDS-PAGE and Western Blot of PotD  </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew8-5"></span>
+                <div id="secollapsiblew8-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To run SDS-PAGE of the elutions obtained on Friday after SEC.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> SDS-PAGE</a></p>
+              <p><b>Results:</b><a data-toggle="modal" data-target="#sbp-figure7-modal" class="journal-protocol"> Figure 7 </a> </p>
+<p><b>Next Steps:</b>The same samples will be Western Blotted on Monday to confirm that the bands oberved are PotD.</p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 9 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 27/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew9"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">PotD was successfully purified and characterized this week. SBP was still unobservable in induced 150ml cultures so 3L cultures containing pQE80-L-<i>SBP</i> were set up. </p>
+         </div>
+         <div id="secollapsiblew9" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">27/7: Overnight Cultures for <i>SBP</i> Expression Assays and SDS-PAGE and Western Blot of PotD</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew9-1"></span>
+                <div id="secollapsiblew9-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up a 150ml overnight culture containing pQE80-L-<i>SBP</i> for sub-culturing tomorrow and to run an SDS-PAGE and Western Blot the PotD samples from Friday.</p>
+                  <p><b>Protocols Used: </b><i>SBP</i>: <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> Note: 150ml of LB was used and 150µl of each antibiotic. <i>PotD</i>: <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> SDS-PAGE and Western Blotting</a> Note: The membrane was left blocking overnight.</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>150ml of fresh LB will be inoculated tomorrow with 7.5ml of the overnight cultures containing pQE80-L-<i>SBP</i>. The PotD Western Blot will be completed tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">28/7: <i>SBP</i> Expression Assay and Completion of PotD Westen Blot</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew9-2"></span>
+                <div id="secollapsiblew9-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up day cultures using the overnight cultures from yesterday and test for expression of LbpA 6 hours after induction.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Test </a> Note: 150ml of fresh LB was innoculated with 7.5ml of the overnight cultures. Only an uninduced control and an 1mM IPTG induced culture was set up. The samples were not blotted, they were only ran on an SDS gel. </p>
+              <p><b>Results:</b> <i>SBP</i>: Nothing was observable on the SDS gel. <i>PotD</i>: <a data-toggle="modal" data-target="#sbp-figure8-modal" class="journal-protocol"> Figure 8 </a> </p>
+<p><b>Next Steps:</b> Since PotD has now been successful purified, the next step is to attach a fluorescent nanobead to the protein. Hopefully, this will be attempted in the coming weeks. Even though SBP was not visible on the SDS gel, 3L cultures will be set up this week to try and harvest the protein since the culture size may be too small. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">29/7: Overnight Cultures for 3L Culture</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew9-3"></span>
+                <div id="secollapsiblew9-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures containing pQE80-L-<i>SBP</i> for the 3L day cultures that will be grown tomorrow.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> Note: One 150ml overnight culture was set up using 150µl of each antibiotic. </p>
+              <p><b>Results:</b> N/A   </p>
+<p><b>Next Steps:</b> The 3L culture will be set up tomorrow allowed to grow to OD<sub>600</sub> of 0.6-1 and then <i>SBP</i> expression induced with 1mM IPTG and grown for 6 hours.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">30/7: 3L Culture for Purification of SBP</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew9-4"></span>
+                <div id="secollapsiblew9-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up 3L day cultures for purifying SBP.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: The procedure was stopped at Step 6 and the pellets were frozen at -20<sup>o</sup>C. </p>
+              <p><b>Results:</b> N/A   </p>
+<p><b>Next Steps:</b> Protein purification will be continued on Monday.</p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+  <!-- WEEK 10 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 3/8/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew10"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">It seems that SBP has been successfully purified and characterized. However, there was still a lot of contamination in the samples after size eclusion chromatography was performed. .</p>
+         </div>
+         <div id="secollapsiblew10" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">3/8: Continuation of SBP Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew10-1"></span>
+                <div id="secollapsiblew10-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue with purification of SBP.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: The protocol was continued from Step 7</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>Size exclusion chromatography will be carried out tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">4/8: Continuation of SBP Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew10-2"></span>
+                <div id="secollapsiblew10-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue purification of SBP.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a>Note: The protocol was continued from Step 14. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The fractions corresponding to the peak observed during SEC, will be ran on an SDS gel and Western Blotted tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">6/8: Western Blot of SBP</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew10-3"></span>
+                <div id="secollapsiblew10-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To perform a Western Blot on the samples obtained from Size Exclusion Chromatography yesterday..</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blot </a></p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#sbp-figure9-modal" class="journal-protocol"> Figure 9 </a> </p>
+<p><b>Next Steps: There was still some contamination in the gel filtrated samples so this will have to be removed before a nanobead can be attached to the protein. </b> </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+  <!-- WEEK 11 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 10/8/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew10"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Interactions between PotD and spermidine were analysed via tryptophan fluorescence this week.</p>
+         </div>
+         <div id="secollapsiblew11" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">10/8: Tryptophan Fluorescence</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew11-1"></span>
+                <div id="secollapsiblew11-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To analyse the interaction of PotD and spermidine using tryptophan fluorescence. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#tf-modal" class="journal-protocol"> Tryptophan Fluorescence</a> </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b></p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+         </div>
+       </div>
+   </div>
+</div>
+<!-- BLOOD-->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <span id="biospray-title">Blood</span>
+          <span type="button" class="plusminus-toggleable box-btn glyphicon glyphicon glyphicon-plus-sign" data-toggle="collapse" data-target="#blcollapsible"></span>
+           <div id="blcollapsible" class="collapse biospray-content">
+<!-- WEEK 1 -->
+  <div class="row">
+     <div class="box">
+        <div class="labtitle">Week Beginning 10/6/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew1"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>hHBA</i> and <i>hHBB</i> into the biobrick vector, pSB1C3, was performed this week.</p>
+         </div>
+         <div id="blcollapsiblew1" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">10/6: Restriction Digests of <i>hHBA</i> and <i>hHBB</i> from IDT gBlocks and Ligation into pSB1C3</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew1-1"></span>
+                <div id="blcollapsiblew1-1" class="collapse box-content">
+                  <p><b>Aim of Experiment:</b> To digest <i>hHBA</i> and <i>hHBB</i> from IDT gBlocks using EcoRI and PstI for ligation into pSB1C3.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests</a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a></p>
+         <p><b>Results: </b>N/A</p>
+                  <p><b>Next Steps:</b> The pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> ligations will be transformed into JM110 <i>E.coli</i>. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">11/6: Transformations of pSB1C3-hHBA and pSB1C3-hHBB Ligations into JM110 <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew1-2"></span>
+                <div id="blcollapsiblew1-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform the pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> ligations into JM1110 <i>E.coli</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol">Transformations</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> If transformations are successful, overnight cultures will be set up.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+  </div>
+<!-- WEEK 2 -->
+  <div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 15/6/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew2"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">The sequence for pSB1C3-<i>hHBB</i> was confirmed this week so cloning of <i>hHBB</i> into the two hybrid system vector pUT18 was started. Sequencing of pSB1C3-<i>hHBA</i> was incorrect so another miniprep was sent for sequencing.</p>
+         </div>
+         <div id="blcollapsiblew2" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">15/6: Overnight Cultures of JM110 <i>E.coli</i> containing pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew2-1"></span>
+                <div id="blcollapsiblew2-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of JM110 <i>E.coli</i> containing pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> from the transformations performed on Tuesday.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> Overnight cultures will be miniprepped tomorrow and a test restriction digest will be performed to check for the presence of the inserts.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">16/6: Plasmid Purification of the JM110 <i>E.coli</i> Overnight Cultures containing pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew2-2"></span>
+                <div id="blcollapsiblew2-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the JM110 <i>E.coli</i> overnight cultures containing pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> and perform a pre-sequence digest check.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol">Plasmid Purification (QIAprep® Spin Miniprep Kit) </a> <a data-toggle="modal" data-target="#restrictriondigest-modal" class="journal-protocol">Restriction Digests</a> </p>
+         <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure1-modal" class="journal-protocol"> Figure 1 </a>
+       <p><b>Next Steps:</b> The gel indicates that <i>hHBA</i> and <i>hHBB</i> have inserted successfully into pSB1C3 so samples of the minipreps will be sent for sequencing to confirm this.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">17/6: Amplification of <i>hHBA</i> and <i>hHBB</i> for Cloning into the Two Hybrid System Vectors</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew2-3"></span>
+                <div id="blcollapsiblew2-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplify <i>hHBA</i> and <i>hHBB for Cloning into the Two Hybrid System Vectors pT25 and pUT18, respectively. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol">PCR</a></p>
+         <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure2-modal" class="journal-protocol"> Figure 2 </a>
+       <p><b>Next Steps:</b> Sequencing of pSB1c3-<i>hHBA</i> and pSB1c3-<i>hHBB</i> showed that pSB1c3-<i>hHBA</i> had a 1bp deletion so one of the other minipreps was sent for sequencing. pSB1c3-<i>hHBB</i>, however, was successful so tHe band for <i>hHBB</i> will be gel extracted and digested tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">18/6: Gel Extraction and Restriction Digests <i>hHBB</i> for Cloning into pUT18 </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew2-4"></span>
+                <div id="blcollapsiblew2-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To gel extract and digest <i>hHBB</i> with BamHI and EcoRI.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol">Restriction Digests</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> Ligations of <i>hHBB</i> will be set up tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">19/6: Ligations of <i>hHBB</i> into pUT18</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew2-4"></span>
+                <div id="blcollapsiblew2-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up ligations of pUT18-<i>hHBB</i>.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol">Ligations</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> Ligations will be left over the weekend and transformed into BTH101 <i>E.coli</i>. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 3 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 22/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew3"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content"> Cloning of <i>hHBA</i> and <i>hHBB</i> into pT25 and pUT18, respectively, was continued this week.</p>
+         </div>
+         <div id="blcollapsiblew3" class="collapse week-content">
+     <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">22/6: Transformations of pUT18-<i>hHBB</i> into BTH101 <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew3-1"></span>
+                <div id="blcollapsiblew3-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform pUT18-<i>hHBB</i> into BTH101 <i>E.coli</i>.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations</a></p>
+                  <p><b>Results:</b>N/A </p>
+<p><b>Next Steps:</b>If transformations are successful, overnight cultures will be set up tomorrow. Sequencing showed the sequence for pSB1C3-<i>hHBA</i> was incorrect again, a third miniprep will be sent for sequencing.</p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">23/6: Overnight Cultures of BTH101 <i>E.coli</i> containing pUT18-<i>hHBB</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew3-2"></span>
+                <div id="blcollapsiblew3-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of BTH101 <i>E.coli</i> containing pUT18-<i>hHBB</i> from the transformations carried out yesterday.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures</a></p>
+                  <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The overnight cultures will be miniprepped and sent for sequencing tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">24/6: Plasmid Purification of the BTH101 <i>E.coli</i> Overnight Cultures containing pUT18-<i>hHBB</i></i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew3-3"></span>
+                <div id="blcollapsiblew3-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the BTH101 <i>E.coli</i> Overnight Cultures containing pUT18-<i>hHBB</i>.</p>
+                  <p><b>Protocols Used: </b></p><a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a></p>
+                  <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> The miniprep with the highest concentration will be sent for sequencing to confirm presence of insert. Sequencing for pSB1C3-<i>hHBA</i> came back correct so another PCR of <i>hHBA</i> will be set up tomorrow for cloning into pT25. </p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">25/6: Amplification of <i>hHBA</i> for Cloning into pT25</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3-4"></span>
+                <div id="secollapsiblew3-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplify <i>hHBA</i> for Cloning into pT25</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#pcr-modal" class="journal-protocol"> PCR</a></p>
+                  <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure3-modal" class="journal-protocol"> Figure 3 </a> </p>
+<p><b>Next Steps:</b> The PCR product digested and ligated into pT25 tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">26/6: Restriction Digests and Ligations of <i>hHBA</i> for Cloning into pT25</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew3-5"></span>
+                <div id="secollapsiblew3-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest the <i>hHBA</i> PCR product with BamHI and KpnI and set up ligations with pT25.</p>
+                  <p><b>Protocols Used: </b><a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests</a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a></p>
+                  <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Ligations will be left over the weekend and transformed into BTH101 <i>E.coli</i> on Monday. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 4 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 29/6/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew4"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>hHBA</i> into pT25 was completed this week. Sequencing confirmed that <i>hHBB</i> has been successfully inserted into pUT18. Cloning of both <i>hHBA</i> and <i>hHBB</i>, individually, into the high expression vector pQE80-L was started this week.</p>
+         </div>
+         <div id="blcollapsiblew4" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">29/6: Transformation of pT25-<i>hHBA</i> Ligations into BTH101 <i>E.coli</i>/span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew4-1"></span>
+                <div id="blcollapsiblew4-1" class="collapse box-content">
+                  <p><b>Aim of Experiment:</b> To transform pT25-<i>hHBA</i> into  BTH101 <i>E.coli</i>.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations</a>></p>
+         <p><b>Results: </b>N/A</p>
+                  <p><b>Next Steps:</b> Overnight cultures will be set up tomorrow for the pT25-<i>hHBA</i> transformations if successful. Sequencing confirmed <i>hHBB</i> has been successfully inserted into pUT18. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">30/6: Overnight Cultures of MC1061 <i>E.coli</i> containing pUT18-<i>hHBA</i> and Amplification of <i>hHBA</i> and <i>hHBB</i> for Cloning into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew4-2"></span>
+                <div id="blcollapsiblew4-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of MC1061 <i>E.coli</i> containing pT25-<i>hHBA</i>. To amplify <i>hHBA</i>.and <i>hHBB</i> for cloning into pQE80-L.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a>  <a data-toggle="modal" data-target="#pcr-modal" class="journal-protocol"> Transformations </a><a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> PCR </a></p>
+         <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure4-modal" class="journal-protocol"> Figure 4 </a> </p>
+       <p><b>Next Steps:</b> The BTH101 <i>E.coli</i> overnight cultures containing pT25-<i>hHBB</i> will be miniiprepped tomorrow. The PCR products will be digested and ligate dinto pQE80-L tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">1/7: Plasmid Purification of the Overnight Cultures of BTH101 <i>E.coli</i> containing pT25-<i>hHBA</i> and Restriction Digests and Ligations of <i>hHBA</i> and <i>hHBB</i> into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew4-3"></span>
+                <div id="blcollapsiblew4-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the overnight cultures of BTH101 <i>E.coli</i> containing pT25-<i>hHBA</i>. To digest <i>hHBA</i> and <i>hHBB</i> with BamHI and KpnI and set up ligations with pQE80-L.</p>
+                  <p><b>Protocols Used:</b><a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations </a> </p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The miniprep with the highest concentration for pT25-<i>hHBA</i> will be sent for sequencing. The pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i> will be transformed into M15[pREP4] <i>E.coli</i> tomrorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">2/7: Transformations of pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i> Ligations into M15[pREP4] <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew4-4"></span>
+                <div id="blcollapsiblew4-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform the pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i> ligations into M15[pREP4] <i>E.coli</i>.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> If transformations are successful, overnight cultures will be set up on Monday. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 5 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 6/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew5"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>hHBA</i> and <i>hHBB</i> into pQE80-L was completed this week. Cloning of <i>Haptoglobin (hHBN) </i> was also started this week.</p>
+         </div>
+         <div id="blcollapsiblew5" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">6/7: Overnight Cultures of m15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew5-1"></span>
+                <div id="blcollapsiblew5-1" class="collapse box-content">
+                  <p><b>Aim of Experiment:</b> To set up overnight cultures of BTH101 <i>E.coli</i> containing pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> </p>
+         <p><b>Results: </b> N/A </p>
+      <p><b>Next Steps:</b>The overnight cultures will be miniprepped tomorrow. Sequencing for pT25-<i>hHBA</i> was incorrect so cloning will be repeated from the restriction digest stage.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">7/7: Plasmid Purification of m15[pREP4] <i>E.coli</i> Overnight Cultures containing pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew5-2"></span>
+                <div id="blcollapsiblew5-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up miniprep the BTH101 <i>E.coli</i> Overnight Cultures containing pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol">  Plasmid Purification (QIAprep® Spin Miniprep Kit) </a></p>
+         <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure5-modal" class="journal-protocol"> Figure 5</a> </p>
+       <p><b>Next Steps:</b> The pre-sequence digest indicates that both <i>hHBA</i> and <i>hHBB</i> have been inserted into pQE80-L successfully. The minipreps with the highest concentration will be sent for sequencing. The sequence for <i>Haptoglobin (hHBN)</i> arrived in an IDT plasmid, work will be be started on this tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">8/7: Transformation of IDT Plasmid-<i>hHBN</i> into MC1061 <i>E.coli</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew5-3"></span>
+                <div id="blcollapsiblew5-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform the IDT Plasmid-<i>hHBN</i> into MC1061 <i>E.coli</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations</a> </p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> Overnight cultures will be set up tomorrow. </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">9/7: Overnight Cultures of MC1061 <i>E.coli</i> containing IDT Plasmid-<i>hHBN</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew5-4"></span>
+                <div id="blcollapsiblew5-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of MC1061 <i>E.coli</i> containing IDT Plasmid-<i>hHBN</i>. </p>
+         <p><b>Results:</b> N/A</p>
+       <p><b>Next Steps:</b> The overnight cultures will be miniprepped tomorrow and <i>hHBN</i> will be amplified for cloning into pSB1C3. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">10/7: Plasmid Purification of IDT Plasmid-<i>hHBN</i> and Digestion of <i>hHBN</i> for Cloning into pSB1C3</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew5-5"></span>
+                <div id="blcollapsiblew5-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the MC1061 <i>E.coli</i> containing IDT Plasmid-<i>hHBN</i> and to digest <i>hHBN</i> out of the plasmid using EcoRI and PstI.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests</a> </p>
+         <p><b>Results:</b> N/A</p>
+       <p><b>Next Steps:</b> <i>hHBN</i> will be ligated into pSB1C3 on Monday. Sequencing also confirmed <i>hHBA</i> and <i>hHBB</i> have been successfully inserted into pQE80-L.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 6 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 13/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew6"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">hHBB was successfully characterized this week so protein purification was started. Cloning of <i>hHBN</i> into pSB1C3 was also completed.</p>
+         </div>
+         <div id="blcollapsiblew6" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">13/7: Ligations of <i>hHBN</i> into pSB1C3 and Overnight Cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew6-1"></span>
+                <div id="blcollapsiblew6-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To ligate <i>hHBN</i> into pSB1C3. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures</a> </p>
+              <p><b>Results:</b>N/A</p>
+<p><b>Next Steps:</b> The pSB1C3-<i>hHBN</i> ligations will be transformed into MC1061 <i>E.coli</i> tomorrow. The overnight cultures will be subcultured tomorrow and induced with IPTG to test protein expression levels. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">14/7: Transformation of pSB1C3-<i>hHBN</i> into MC1061 <i>E.coli</i> and Optmiization of <i>hHBA</i> and <i>hHBB</i> Expression </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew6-2"></span>
+                <div id="blcollapsiblew6-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform pSB1C3-<i>hHBN</i> into MC1061 <i>E.coli</i> and set up expression optimization tests for <i>hHBA</i> and <i>hHBB</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Protein Expression Optimization </a> </p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure6-modal" class="journal-protocol"> Figure 6 </a></p>
+<p><b>Next Steps:</b> If transformations are successful, overnight cultures will be set up tomorrow. Western Blotting will be continued tomorrow for <i>hHBA</i> and <i>hHBB</i>. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">15/7: Overnight Cultures of MC1061 <i>E.coli</i> containing pSB1C3-<i>hHBN</i> and Western Blotting of <i>hHBA</i> and <i>hHBB</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew6-3"></span>
+                <div id="blcollapsiblew6-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight culture of MC1061 <i>E.coli</i> containing pSB1C3-<i>hHBN</i> and finish Western Blotting of <i>hHBA</i> and <i>hHBB</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blotting</a> </p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure7-modal" class="journal-protocol"> Figure 7 </a> </p>
+<p><b>Next Steps:</b> The blot shows that characterization of hHBB has been successful. Unfortunately, nothing was visible on the blot for hHBA. 3L cultures for purification of hHBB will be set up this week. Characterization of hHBA may be reattempted. The overnight cultures will be miniprepped tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">16/7: Plasmid Purification of pSB1C3-<i>hHBN</i> and Overnight Cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBB</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew6-4"></span>
+                <div id="blcollapsiblew6-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the MC1061 <i>E.coli</i> overnight cultures containing pSB1C3-<i>hHBN</i> and set up overnight cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBB</i> for 3L cultures. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The miniprep with the highest concentration will be sent for sequencing. The overnight cultures will be used to set up 3L day cultures tomorrow for purrification of hHBB. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">17/7: 3L Cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBB</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew6-5"></span>
+                <div id="blcollapsiblew6-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up 3L day cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBB</i> for purification of hHBB. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: The protocol was stopped at Step 6. </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> Purification will be continued on Monday.</p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 7 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 20/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew7"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">hHBB was successfully purified this week and cloning of <i>hHBN</i> into pQE80-L was completed. </p>
+         </div>
+         <div id="blcollapsiblew7" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">20/7: Amplification of <i>hHBN</i> for Cloning into pQE80-L and Purification of hHBB</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew7-1"></span>
+                <div id="blcollapsiblew7-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplify <i>hHBN</i> for cloning into pQE80-L and continuation of hHBB purification.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pcr-modal" class="journal-protocol"> PCR </a> <b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: Protocol was continued from Step 6 <a data-toggle="modal" data-target="#pcr-modal" class="journal-protocol"> PCR</a></p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure8-modal" class="journal-protocol"> Figure 8</a> </p>
+<p><b>Next Steps:</b> The fractions from the affinity chromatography will be consolidated and concentrated tomorrow. Size exclusion chromatography will also be performed. The <i>hHBN</i> PCR product will be digested and ligated into pQE80-L tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">21/7: Size Exclusion Chromatography of hHBB and Restriction Digests and Ligations of <i>hHBN</i> into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew7-2"></span>
+                <div id="blcollapsiblew7-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To consolidate, concentrate and perform size exclusion chromatography on the hHBB samples. To digest the <i>hHBN</i> PCR product with BamHI and KpnI and subsequently ligate it into pQE80-L. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> SDS </a> Note: An SDS gel was ran of the SEC fractions which corresponded to the observed peak. Western blotting was stopped at the blocking stage. <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations </a></p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure9-modal" class="journal-protocol"> Figure 9 </a> </p>
+<p><b>Next Steps:</b> Western blotting will be completed tomorrow and will confirm if the bands observed on the SDS gel are indeed hHBB. The pQE80-L<i>hHBN</i> ligations will be transformed into M15[pREP4] <i>E.coli</i> tomorrrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">22/7: Western Blot of hHBB and Transformations of m15[pREP4] <i>E.coli</i> with pQE80-L<i>hHBN</i> Ligations</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew7-3"></span>
+                <div id="blcollapsiblew7-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To complete the Western blot of hHBB and transform pQE80-L<i>hHBN</i> into m15[pREP4] <i>E.coli</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blotting </a> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a>  </p>
+              <p><b>Results:</b><a data-toggle="modal" data-target="#bl-figure10-modal" class="journal-protocol"> Figure 10 </a></p>
+<p><b>Next Steps:</b> The blot shows hHBB has been successfully purified. If the transformations are successful, overnight cultures will be set up tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">23/7: Overnight Cultures of m15[pREP4] <i>E.coli</i> containing pQE80-L<i>hHBN</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew7-4"></span>
+                <div id="blcollapsiblew7-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of m15[pREP4] <i>E.coli</i> containing pQE80-L<i>hHBN</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> The overnight cultures will be miniprepped tomorrow.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">24/7: Plasmid Purification of Overnight Cultures of m15[pREP4] <i>E.coli</i> containing pQE80-L<i>hHBN</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew7-5"></span>
+                <div id="blcollapsiblew7-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep m15[pREP4] <i>E.coli</i> overnight cultures containing pQE80-L<i>hHBN</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> The miniprep with the highest concentration will be sent for sequencing. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 8 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 27/7/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew8"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content"><i>hHBN</i> was successfully cloned into pQE80-L and cloning started for insertion into pT25.</p>
+         </div>
+         <div id="blcollapsiblew8" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">27/7: Restriction Digests and Ligations of <i>hHBN</i> into pQE80-L</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#secollapsiblew8-1"></span>
+                <div id="secollapsiblew8-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest <i>hHBN</i> again with BamHI and KpnI and set up ligations with pQE80-L.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Resriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations </a> </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> Ligations will be transformed into M15[pREP4] <i>E.coli</i> tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">28/7: Transformation of M15[pREP4] <i>E.coli</i> with pQE80-L-<i>hHBN</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew8-2"></span>
+                <div id="blcollapsiblew8-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform M15[pREP4] <i>E.coli</i> with pQE80-L-<i>hHBN</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a></p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> If transformations are successful, overnight cultures will be set up tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">29/7: Overnight Cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBN</i>  </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew8-3"></span>
+                <div id="blcollapsiblew8-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBN</i> </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The overnight cultures will be miniprepped tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">30/7: Plasmid Purification of M15[pREP4] <i>E.coli</i> Overnight Cultures containing pQE80-L-<i>hHBN</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew8-4"></span>
+                <div id="blcollapsiblew8-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the M15[pREP4] <i>E.coli</i> Overnight Cultures containing pQE80-L-<i>hHBN</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>The miniprep with the highest concentration will be sent for sequencing. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">31/7: Amplification of <i>hHBN</i> for Cloning into pT25</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew8-5"></span>
+                <div id="blcollapsiblew8-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplfy <i>hHBN</i> for cloning into the two hybrid vector pT25.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pcr-modal" class="journal-protocol"> PCR</a></p>
+              <p><b>Results:</b><a data-toggle="modal" data-target="#bl-figure11-modal" class="journal-protocol"> Figure 11 </a> </p>
+<p><b>Next Steps:</b>The PCR product will be digested and ligated into pT25 on Monday.</p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 9 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 3/8/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew9"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>hHBN</i> into pT25 was completed this week but subsequent sequencing showed that it had not inserted successfully. </p>
+         </div>
+         <div id="blcollapsiblew9" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">3/8: Restriction Digests and Ligations of <i>hHBN</i> for Cloning into pT25</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew9-1"></span>
+                <div id="blcollapsiblew9-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest the <i>hHBN</i> PCR product with BamHI and KpnI and set up ligations with the pT25 vector.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol"> Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol"> Ligations</a> </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>The pT25-<i>hHBN</i> ligations will be transformed into MG1655 <i>E.coli</i> tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">4/8: Transformations of  pT25-<i>hHBN</i> into MG1655 <i>E.coli</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew9-2"></span>
+                <div id="blcollapsiblew9-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up transformations of pT25-<i>hHBN</i> into MG1655 <i>E.coli</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol"> Transformations </a> </p>
+              <p><b>Results:</b> N/A</p>
+<p><b>Next Steps:</b> If transformations are successful, overnight cultures will be set up tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">5/8: Overnight Cultures of MG1655 <i>E.coli</i> containing pT25-<i>hHBN</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew9-3"></span>
+                <div id="blcollapsiblew9-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of MG1655 <i>E.coli</i> containing pT25-<i>hHBN</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a> </p>
+              <p><b>Results:</b> N/A   </p>
+<p><b>Next Steps:</b> The overnight cultures will be miniprepped tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">6/8: Plasmid Purification of MG1655 <i>E.coli</i> Overnight Cultures containing pT25-<i>hHBN</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew9-4"></span>
+                <div id="blcollapsiblew9-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the  MG1655 <i>E.coli</i> overnight cultures containing pT25-<i>hHBN</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Plasmid Purification (QIAprep® Spin Miniprep Kit)</a> </p>
+              <p><b>Results:</b> N/A   </p>
+<p><b>Next Steps:</b> The miniprep with the highest concentration will be sent for sequencing. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+  <!-- WEEK 10 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 10/8/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew10"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Overexpression assays of <i>hHBN</i> were set up this week and purification of the protein was begun. </p>
+         </div>
+         <div id="blcollapsiblew10" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">10/8: Overnight Cultures for <i>hHBN</i> Overexpression Assays</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#sblcollapsiblew10-1"></span>
+                <div id="blcollapsiblew10-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBN</i>. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures </a></p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>The overnight cultures will be used to set up day cultures that will be induced with different concentrations of IPTG to assay <i>hHBN</i> expression levels.</p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">11/8: Overexpression Assay of pQE80-L-<i>hHBN</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew10-2"></span>
+                <div id="blcollapsiblew10-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To assay <i>hHBN</i> expression levels when induced with a range of IPTG concentrations. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Overexpression Assay </a>Note: Day cultures were grown to an OD<sub>600</sub> of 0.6 and then induced with 0.25 mM, 0.5 mM, 0.75 mM and 1 mM IPTG and allowed to grow for 6 hours. </p>
+              <p><b>Results:</b> SDS gel </p>
+<p><b>Next Steps:</b> The gel indicates that <i>hHBN</i> is not being overexpressed in any of the cultures. Further optimization experiments will be set up to try and overexpress <i>hHBN</i>. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">12/8: Overnight Cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBN</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew10-3"></span>
+                <div id="blcollapsiblew10-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBN</i> which will be used to set up a 3 L day culture tomorrow.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> Overnight Cultures</a> Note: A 150 ml overnight culture was set up. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The overnight cultures will be used to set up 3 L day cultures tomorrow.  </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">13/8: 3L Day Cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBN</i></span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew10-4"></span>
+                <div id="blcollapsiblew10-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up 3L day cultures of M15[pREP4] <i>E.coli</i> containing pQE80-L-<i>hHBN</i> using the overnight culture set up yesterday.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> Note: The cultures were grown to OD<sub>600</sub>=0.9, then induced with 1 mM IPTG and allowed to grow for a further 8 hours. A 1 ml sample was taken before the cultures were spun down. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> Protein purification will be continued next week. </p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+  <!-- WEEK 11 -->
+<div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 17/8/15</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew11"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Purification of haptoglobin was completed this week and confirmed via Western Blotting. </p>
+         </div>
+         <div id="blcollapsiblew11" class="collapse week-content">
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">17/8: hHBN Purification</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew11-1"></span>
+                <div id="blcollapsiblew11-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue purification of hHBN by performing affinity chromatography. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification </a> Note: The protocol was stopped at Step 13.</p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b>The samples corresponding to the peak observed in the chromatogram will be combined and purified using SEC tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">18/8: hHBN Purification </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew11-2"></span>
+                <div id="blcollapsiblew11-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To continue purification of hHBN by performing size exclusion chromatography. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#pp-modal" class="journal-protocol"> Protein Purification</a> </p>
+              <p><b>Results:</b> </p>
+<p><b>Next Steps</b>:The gel shows bands corresponding to 45kDa, the expected size of hHBN. This will be checked via Western Blotting to ensure it is the correct protein. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">19/8: Western Blotting of Purified hHBN</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew11-3"></span>
+                <div id="blcollapsiblew11-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To perform Western Blotting in order to verify the protein that was purified is indeed hHBN. </p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blotting </a> Note: The membrane was left blocking in milk overnight. </p>
+              <p><b>Results:</b> N/A </p>
+<p><b>Next Steps:</b> The blot will be completed tomorrow. </p>
+</div>
+        </div>
+        </div>
+      </div>
+<div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">20/8: Western Blotting of Purified hHBN</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew11-4"></span>
+                <div id="blcollapsiblew11-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To complete Western Blotting of hHBN.</p>
+                  <p><b>Protocols Used: </b> <a data-toggle="modal" data-target="#sds-modal" class="journal-protocol"> Western Blotting</a> The protocol was continued from Step 12.</p>
+              <p><b>Results:</b> <a data-toggle="modal" data-target="#bl-figure14-modal" class="journal-protocol"> Figure 14</a> </p>
+<p><b>Next Steps:</b> The blot indicates that hHBN has been successfully purified.</p>
+</div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 12 -->
+  <div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 24/8/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew12"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>hHBA</i> into pT25 was carried out this week. </p>
+         </div>
+         <div id="blcollapsiblew12" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">24/8: Amplification of <i>hHBA</i> for Cloning into pT25</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew12-1"></span>
+                <div id="blcollapsiblew12-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplify <i>hHBA</i> for subsequent cloning into the two hybrid vector pT25.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#pcr-modal" class="journal-protocol">PCR</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The gel extracted PCR product will be digested and ligated into pT25 tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">25/8: Restriction Digests of <i>hHBA</i> and Ligations into pT25 </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew12-2"></span>
+                <div id="blcollapsiblew12-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest the <i>hHBA</i> with BamHI and KpnI and set up ligations with pT25. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol">Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol">Ligations</a> </p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b>The ligations will be left overnight and transformed into MG1065∆cya <i>E.coli</i> tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">26/8: Transformations of pT25-<i>hHBA</i> into MG1065∆cya <i>E.coli</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew12-3"></span>
+                <div id="blcollapsiblew12-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform the pT25-<i>hHBA</i> ligations into MG1065∆cya <i>E.coli</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol">Transformations </a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> If transformations are successful, colonies will be pick for setting up overnight cultures tomorrow.  </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">27/8: Overnight Cultures of MG1065∆cya <i>E.coli</i> containing pT25-<i>hHBA</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew12-4"></span>
+                <div id="blcollapsiblew12-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of MG1065∆cya <i>E.coli</i> containing pT25-<i>hHBA</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The overnight cultures will be miniprepped tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">28/8: Plasmid Purification of MG1065∆cya <i>E.coli</i> containing pT25-<i>hHBA</i> Overnight Cultures</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew12-5"></span>
+                <div id="blcollapsiblew12-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the  MG1065∆cya <i>E.coli</i> overnight cultures containing pT25-<i>hHBA</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol">Plasmid Purification (QIAprep® Spin Miniprep Kit)</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The miniprep with highest concentration was sent for sequencing to confirm if the insert has been cloned in successfully. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+<!-- WEEK 13 -->
+  <div class="row">
+    <div class="border-week">
+     <div class="box">
+        <div class="labtitle">Week Beginning 7/9/2015</div>
+         <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew13"></span>
+          <div class="box-content">
+          <p class="journal-summary-heading">Summary</p>
+         <p class="journal-content">Cloning of <i>hHBN</i> into pUT18 was carried out this week. </p>
+         </div>
+         <div id="blcollapsiblew13" class="collapse week-content">
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">24/8: Amplification of <i>hHBN</i> for Cloning into pUT18</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew13-1"></span>
+                <div id="blcollapsiblew13-1" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To amplify <i>hHBN</i> for subsequent cloning into the two hybrid vector pUT18.</p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#pcr-modal" class="journal-protocol">PCR</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The gel extracted PCR product will be digested and ligated into pT25 tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">25/8: Restriction Digests of <i>hHBN</i> and Ligations into pUT18 </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew13-2"></span>
+                <div id="blcollapsiblew13-2" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To digest the <i>hHBN</i> with BamHI and KpnI and set up ligations with pUT18. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#restrictiondigest-modal" class="journal-protocol">Restriction Digests </a> <a data-toggle="modal" data-target="#ligation-modal" class="journal-protocol">Ligations</a> </p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b>The ligations will be left overnight and transformed into MG1065∆cya <i>E.coli</i> tomorrow. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">26/8: Transformations of pUT18-<i>hHBN</i> into MG1065∆cya <i>E.coli</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew13-3"></span>
+                <div id="blcollapsiblew13-3" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To transform the pUT18-<i>hHBN</i> ligations into MG1065∆cya <i>E.coli</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol">Transformations </a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> If transformations are successful, colonies will be pick for setting up overnight cultures tomorrow.  </p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">27/8: Overnight Cultures of MG1065∆cya <i>E.coli</i> containing pUT18-<i>hHBN</i> </span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew13-4"></span>
+                <div id="blcollapsiblew13-4" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To set up overnight cultures of MG1065∆cya <i>E.coli</i> containing pUT18-<i>hHBN</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The overnight cultures will be miniprepped tomorrow.</p>
+                </div>
+        </div>
+        </div>
+      </div>
+      <div class="row">
+       <div class="border-day">
+        <div class="box">
+            <span class="box-title">28/8: Plasmid Purification of MG1065∆cya <i>E.coli</i> containing pUT18-<i>hHBN</i> Overnight Cultures</span>
+              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#blcollapsiblew13-5"></span>
+                <div id="blcollapsiblew13-5" class="collapse box-content">
+                  <p><b>Aim of experiment:</b> To miniprep the  MG1065∆cya <i>E.coli</i> overnight cultures containing pUT18-<i>hHBN</i>. </p>
+                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol">Plasmid Purification (QIAprep® Spin Miniprep Kit)</a></p>
+         <p><b>Results:</b> N/A
+       <p><b>Next Steps:</b> The miniprep with highest concentration was sent for sequencing to confirm if the insert has been cloned in successfully. </p>
+                </div>
+        </div>
+        </div>
+      </div>
+         </div>
+     </div>
+   </div>
+  </div>
+         </div>
+       </div>
+   </div>
+</div>
+<div class="modal-footer">
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+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure1-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
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+        <div class="modal-title" id="myModalLabel">Figure 1: <b>LbpA PCR. Lane 1: LbpA PCR Lane 2: 1kb Ladder and Marker. </b> The image shows the gel ran of the LbpA PCR product. The expected size of the fragment was 2832 bp which corresponds to the observed band on the gel.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/8/85/Dundee15-LbpAPCR-pSB1C3.jpg" style="width:400px;height:auto"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure2-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 2: LbpA PCR. Lane 1: PCR Product, Lane 2: 1kb Ladder and Marker.</b> The image shows the gel ran of the LbpA PCR product. The expected size of the fragment was 2832 bp which corresponds to the observed band on the gel.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/d/d4/LbpA_PCR_pqe80_.jpg" style="width:400px;height:auto"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure3-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 3: LbpA PCR for Cloning into pQE80-L. Lane 1: LbpA PCR Product, Lane 2: 1kb Ladder and Marker</b> The image shows the gel ran of the LbpA PCR product. The expected size of the fragment was 2832 bp which corresponds to the observed band on the gel.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/d/d4/LbpA_PCR_pqe80_.jpg" style="width:400px;height:auto"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure4-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 4: LbpA Culture Samples. Lane 1: Sample from uninduced culture. Lane 2: Sample from culture induced with 1mM IPTG.</b> It seems that on comparison of the two cultures, that inducing expression of LbpA causes the cells to die given the significant reduction of protein levels visible on the gel. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/a/a6/LbpA_sds_030715.png" style="width:150px;height:auto"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure5-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 5: OD<sub>600</sub> Readings from Uninduced Culture and Induced Cultures (0.5 mM, 1 mM and 2 mM IPTG).</b> The table shows that after being induced with IPTG the cultures stop growing since their OD<sub>600</sub> readings are notably lower than that of the uninduced control. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/f/f4/LbpA_table_070715.jpg" style="width:500px;height:auto"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure6-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 6: Growth Curve of Uninduced and Induced Cultures (0.5 mM, 1 mM and 2 mM IPTG)</b> The graph shows that on comparison with the uninduced control, the induced cultures stop growing when LbpA expression is induced. After ~7 hours, it is possible that the emergence of mutants causes the the OD to rise again or the cells have adapted to the production of the foreign protein. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/b/bc/Lbpa_growth_curve_140715.png" style="width:500px;height:auto"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure7-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 7: Growth Curve of Uninduced and Induced Cultures (0.5 mM, 1 mM and 2 mM IPTG)</b> The graph shows that on comparison with the uninduced control, after reaching an OD<sub>600</sub> of 0.6,  the induced cultures stop growing when LbpA starts to be expressed. Similar to the previous growth curve assay, after ~7 hours, it is possible that the emergence of mutants causes the the OD to rise again or the cells have dealt with the production of the foreign protein. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/1/1c/Lbpa_growth_curve_230715.png" style="width:500px;height:auto"> </center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="lbpa-figure8-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 8: SDS Gel of Samples Taken from Uninduced Control and 1 mM IPTG Induced Culture 6 hours After Induction</b> For the induced culture, a faint band just above the 100 kDa marker is observable on the gel which corresponds to the expected size of LbpA.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/f/ff/Lbpa_sds_280715.png" style="width:300px;height:auto"> </center>
+      </div>
+      <div class="modal-footer">
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+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure1-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 1: Gel of <i>SBP</i> PCR Product for Cloning into pQE80-L. Lane 1: <i>SBP</i> PCR Product 1 Lane 2: 1kb Ladder and Marker Lane 3: <i>SBP</i> PCR Product 2</b> Both bands observable on the gel correspond to the expected size of <i>SBP</i> which is ~600 bp.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/e/ef/Sbp_pqe80l_pcr_220615.png" style="width:300px;height:auto"> </center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure2-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 2: SDS Gel of Uninduced and 1 mM IPTG Induced Cultures of PotD and SBP.</b> For PotD, it is clear from the gel that it is being successfully overexpressed since there is an intense band present at the 37 kDa marker which corresponds to the expected size of PotD in the induced culture sample. For SBP, there is no band present which suggests SBP is being overexpressed. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/8/8d/Potd_sbp_sds_020715.png" style="width:300px;height:auto"> </center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure3-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 3: Characterisation of PotD.</b> Single colonies of MC1061 <i>E.coli</i> containing pQE80-L encoding <i>PotD</i> were used to inoculate 5ml of fresh LB growth medium supplemented with ampicillin. Once cultures reached an OD<sub>600</sub> of 0.6, one was induced with 1 mM IPTG and one was left uninduced as a control. They were left to grow for a further three hours, after which a 1ml sample was taken and the cells pelleted. The cells were resuspended in 100µl of Laemmli buffer, 20µl of which was separated by SDS-PAGE and transferred to a nitrocellulose membrane and probed with an anti-His antibody.  </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/0/09/Potd_wb_030715.png" style="width:300px;height:auto"> </center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure4-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 4: SDS Gel of Uninduced and 0.5mM, 1mM and 2mM IPTG Induced Cultures of SBP.</b> It appears that upon induction of <i>SBP</i> with any concentration of IPTG the cells are dying given that there is a significant drop in protein levels observable on the gel.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/0/0e/Sbp_sds_opt_060715.png"> </center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure5-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 5: Growth Curve of Uninduced and Induced Cultures (0.5 mM, 1 mM and 2 mM IPTG)</b> The graph shows that on comparison with the uninduced control, the induced cultures stop growing when SBP expression is induced. After ~7 hours, it is possible that the emergence of mutants causes the the OD to rise again or the cells have adapted to the production of the foreign protein. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/9/95/Sbp_growth_curve_140715.png" style="width:500px;height:auto"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure6-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 6: Growth Curve of Uninduced and Induced Cultures (0.5 mM, 1 mM and 2 mM IPTG)</b> The graph shows that on comparison with the uninduced control, after reaching an OD<sub>600</sub> of 0.6,  the induced cultures stop growing when <i>SBP</i> starts to be expressed. Similar to the previous growth curve assay, after ~7 hours, it is possible that the emergence of mutants causes the the OD to rise again or the cells have adapted to the production of the foreign protein.  </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/d/d4/Sbp_growth_curve_230715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure7-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 7: SDS-PAGE of Fractions Obtained After Size Exclusion Chromatography of PotD</b> 10 µl of each fraction was mixed with 10 µl of 2x Laemmli buffer and loaded onto an SDS gel which was ran for 1 hour at 100 V. The two bands observable on the gel correspond to the expected size of PotD of 38 kDa. Western Blotting will confirm if these proteins are PotD. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/6/63/Potd_sds_240715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure8-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 8: Characterisation of PotD following Purification.</b> 10 µl of each fraction was mixed with 10 µl of 2x Laemmli buffer and loaded onto an SDS gel which was ran for 1 hour at 100V. It was then transferred to a nitrocellulose membrane and probed with an anti-His antibody.  </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/8/83/Potd_wb_280715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="sbp-figure9-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 9: Characterisation of SBP following Purification.</b> 10 µl of each fraction was mixed with 10 µl of 2x Laemmli buffer and loaded onto an SDS gel which was ran for 1 hour at 100V. It was then transferred to a nitrocellulose membrane and probed with an anti-His antibody.  </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/0/07/Sbp_wb_050815.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure1-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 1: pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> (respectively) Pre-Sequencing Digest</b> pSB1C3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> minipreps were digested with EcoRI and PstI to check for presence of each insert. The bands observable on the gel correspond to the expected sizes for the pSB1C3 vector and both inserts.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src=""></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure2-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 2: Amplification of <i>hHBA</i> and <i>hHBB</i> </b> <i>hHBA</i> and <i>hHBB</i> were amplified from the pSB1c3-<i>hHBA</i> and pSB1c3-<i>hHBB</i> minipreps obtained yesterday using primers to allow cloning of the genes into pT25 and pUT18 respectively. Although the band is fainter in the case of <i>hHBA</i>, the gel indicates the PCR has been successful. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/4/4f/Hhba%2Bhhbb_for_pt25%2Bput18.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure3-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 3: Amplification of <i>hHBA</i></b> <i>hHBA</i> was amplified from the pSB1C3-<i>hHBA</i> using primers to allow cloning of the genes into pT25. The gel indicates that the PCR has been successful. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/4/41/PCR_HgA_PT25.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure4-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 4: Amplification of <i>hHBA</i> and <i>hHBB</i> </b> <i>hHBA</i> and <i>hHBB</i> were amplified from the pSB1c3-<i>hHBA</i> and pSB1C3-<i>hHBB</i> minipreps using primers to allow cloning of the genes into pQE80-L. The gel indicates that the PCR has been successful. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/4/41/PCR_HgA_PT25.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure5-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 5: Pre-Sequence Digest of pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i> </b> Both pQE80-L-<i>hHBA</i> and pQE80-L-<i>hHBB</i> minipreps were digested with BamHI and KpnI to check for the presence of the inserts. The gel indicates that both inserts have been successfully cloned into pQE80-L. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/4/4f/Hhba%2Bhhbb_for_pt25%2Bput18.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure6-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b> Overexpression of <i>hHBB</i> and <i>hHBA</i>.</b> Single colonies of M15[pREP4] <i>E.coli</i> containing pQE80-L encoding <i>hHBB</i> and pQE80-L encoding <i>hHBA</i> were used to inoculate 5ml of fresh LB growth medium supplemented with ampicillin and kanamycin. Once cultures reached an OD<sub>600</sub> of 0.6, they were induced with a range of IPTG concentrations. They were left to grow for a further three hours, after which a 1 ml sample was taken and the cells pelleted. The cells were resuspended in 100 µl of 2x Laemmli buffer, 20 µl of which was separated by SDS-PAGE. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/a/aa/Hhbb_hhba_sds_100715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure7-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 7: Characterisation of hHBB.</b> Single colonies of M15[pREP4] <i>E.coli</i> containing pQE80-L encoding <i>hHBB</i> were used to inoculate 5 ml of fresh LB growth medium supplemented with ampicillin and kanamycin. Once cultures reached an OD<sub>600</sub> of 0.6, they were induced with a range of IPTG concentrations. They were left to grow for a further three hours, after which a 1 ml sample was taken and the cells pelleted. The cells were resuspended in 100 µl of Laemmli buffer, 20 µl of which was separated by SDS-PAGE. The cells were resuspended in 100 µl of Laemmli buffer, 20 µl of which was separated by SDS-PAGE and transferred to a nitrocellulose membrane and probed with an anti-His antibody. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/8/87/Hhbb_wb_160715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure8-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 8: Amplification of <i>hHBN</i> for Cloning into pQE80-L </b> PCR product from amplification of <i>hHBN</i> from IDT plasmid using primers for cloninig into pQE80-L. The gel indicates <i>hHBN</i> has been amplified successfully since the observed band corresponds to the expected size of 1200 bp.</div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/c/c1/Hhbn_pcr_pqe80-l_200715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure9-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 9: SDS-PAGE of Fractions Obtained After Size Exclusion Chromatography of hHBB</b> 10 µl of each fraction was mixed with 10 µl of laemmli buffer and loaded onto an SDS gel which was ran for 1 hour at 100V. The four bands observable on the gel correspond to the expected size of PotD of 16 kDa. Western Blotting will confirm if these proteins are hHBB. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/d/d1/Hhbb_sds_purified_2107152.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure10-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 10: Characterisation of hHBB following Purification.</b> 10 µl of each fraction was mixed with 10 µl of 2x Laemmli buffer and loaded onto an SDS gel which was ran for 1 hour at 100V. It was then transferred to a nitrocellulose membrane and probed with an anti-His antibody. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/1/16/Hhbb_wb_purified_220715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure11-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 11: Amplification of <i>hHBN</i> for Cloning into pT25 </b> PCR product from amplification of <i>hHBN</i> from IDT plasmid using primers for cloninig into pT25. The gel indicates <i>hHBN</i> has been amplified successfully since the observed band corresponds to the expected size of 1200 bp. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/1/16/Hhbb_wb_purified_220715.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>
+<div class="modal fade" id="bl-figure14-modal" tabindex="-1" role="dialog" aria-labelledby="myModalLabel">
+  <div class="modal-dialog" role="document">
+    <div class="modal-content">
+      <div class="modal-header">
+        <button type="button" class="close" data-dismiss="modal" aria-label="close" aria-hidden="true" role="dialog" tabindex="-1"><span aria-hidden="true">&times;</span></button>
+        <div class="modal-title" id="myModalLabel"><b>Figure 14: Characterisation of hHBN following Purification.</b> 10 µl of each fraction was mixed with 10 µl of 2x Laemmli buffer and loaded onto an SDS gel which was ran for 1 hour at 100V. It was then transferred to a nitrocellulose membrane and probed with an anti-His antibody. </div>
+      </div>
+      <div class="modal-body">
+       <center><img src="https://static.igem.org/mediawiki/2015/1/1a/Hhbn-pqe80-l_purified_200815.png"></center>
+      </div>
+      <div class="modal-footer">
+        <button type="button" class="btn btn-primary" data-dismiss="modal">Close</button>
+      </div>
+    </div>
+  </div>
+</div>

Difference between revisions of "Team:Dundee/Lab Book/FluID"

Revision as of 11:12, 17 September 2015

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