Team:Leicester/Notebook/Week1
Week 1
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21 June 2015
Go fund me page for the Leicester iGEM team was first created as such: Leicester iGEM Go fund me
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22nd June 2015
09:00-18:00
Start of Labs
Making of agar plates, including ones with chloramphenicol, ready to used the next day to streak bacteria
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23rd June 2015
09:00-18:00
Start of Measurement Study
-Suspension of parts required for each device required in the measurement study. The parts (Promoter and GFP) were then transformed into DH5(alpha) bacteria. This is then placed on agar plates made the day before.
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24th June 2015
09:00-18:00
Measurement Study
Bacteria produced from the streaks from the previous day are then restreaked again
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25 June 2015
09:00-18:00
Measurement Study
The restreaks are taken and placed into liquid culture
Week 2
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29th June 2015
09:00-18:00
Measurement Study
Mini prep of the liquid cultures were made and then nano drops were done
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30th June 2015
1% agarose gels were made to ensure that the products we wanted were present in solution
Week 3
Due to our project supervisor being away we decided that we concentrated on ensuring that our wiki was started and planning our actual project!.
Week 4
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7th July 2015
Human Practises
Have created our synthetic biology survey: Leicester iGEM survey
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10th July 2015
Measurement Study
Use of fluorescence imaging to see the green fluorescence produced from each device. Repeats of each device were produced and placed onto a 96 well plate. First set of rows consist of 200ul of each repeat and next set has 10x dilution.Controls were placed on the bottom row of blank, positive and negative control. Measurements done at about 560nm.
Nano drops were then taken of repeats which consisted of the highest amount of fluorescence. Fluorescence measured using Omega FLUOstar
Week 5
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21st July 2015
Human Practises
Our team T shirts arrived
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22nd July 2015
Measurement Study
Device 3 experiments were done to ensure that the parts we want are in a given construct, we did this by producing gels and had done some minipreps
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23rd July 2015
09:00-18:00
Measurement Study
Liquid cultures produced
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24th July 2015
09:00-18:00
Sponsored Walk
Done a sponsored walk to raise some money for travel and accommodation at the Giant Jamboree in Boston. Walked from Market Harborough to Leicester in the lovely rain :)
Week 6
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27th July 2015
9:00-18:00
Discussion with Professor Ketley about out project in respect to colonisation, etc.
Look into our wiki and the co
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28th July 2015
09:00-18:00
Meeting about funding and trip to Boston. Measurement study aspects done and mini preps of each device.
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29th July 2015
09:00-18:00
Measurement Study
Nano drops to find out concentration of DNA and find out if the right stuff is present
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30th July 2015
09:00-18:00
Measurement Study
Production of 60 repeats for device 3, these were placed on gel and producing plates
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31st July 2015
09:00-18:00
Measurement Study
Liquid cultures produced and Device 3 re-digested
Entered for Jamboree
Week 7
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3rd August 2015
Measurement Study
Biological and technical repeats placed into liquid cultures from plates of device 3
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4th August 2015
09:00-11:00
Measurement Study
Centrifuged and resuspended devices in TGE buffer.
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4th August 2015
11:00-18:00
iGEM project
Taking parts out required for kill switch, which are then transformed into competent bacteria and streaked out onto plates.
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5th August 2015
09:00-18:00
iGEM project
mini preps; nano drop for device 3 and clean up of lab
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6th August 2015
11:00-18:00
iGEM project
kill switch digest
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6th August 2015
09:00-18:00
Measurement Study
96 well plate- using Elizo app however this did not work properly
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7th August 2015
09:00-18:00
Measurement Study
Run gel of digests and ligation- run… to test
Track selection and abstract
Week 8
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10th August 2015
Testing of enzymes to ensure they were sill working by digesting known GFP containing backbone and then placing on 1% agarose gels
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10th August 2015
09:00-18:00
iGEM project
Meeting with Dr Haigh about PCR process which we will use to obtain the nadD, nadE and pncB genes.
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11th August 2015
09:00-18:00
iGEM project
Gels were made of ligated products for kill switch comparing with undigested products
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12th August 2015
09:00-18:00
iGEM project
Production of liquid cultures and growing of non competent bacteria overnight
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13th August 2015
09:00-18:00
iGEM project
Liquid cultures of non competent bacteria
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14th August 2015
09:00-18:00
iGEM project
Production of competent bacteria. Recovering of DNA from plates and allow for transforming.
Week 9
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17th July 2015
iGEM Project
Picking of transformants and then grown overnight
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18th August 2015
iGEM project
Mini preps
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19th August 2015
09:00-18:00
iGEM project
PCRing of nadD, nadE as well as internal pncB; gel extractions and nanodrop
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21st August 2015
09:00-18:00
iGEM project
PCR of pncB exterior; gel extraction and nanodrop
Week 10
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24th August 2015
iGEM project
PCR of pncB exterior; testing pncB ensuring that restriction site is removed; gel extraction; nanodrop; transforming of part BBa_K608002 into cells incubated over night.
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25th August 2015
09:00-18:00
iGEM project
pncB exterior PCR- to make a full gene instead of the 2 components; isolated pSB1C3; Gel extraction; Nanodrop; liquid culture and restreaks of BBa_K608002
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26th August 2015
09:00-18:00
iGEM project
Phusion PCR of nadD, nadE and attempted pncB; pncB PCR again with different dilutions; Gel extraction; Nanodrop; miniprep and nanodrop of BBa_K608002
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27th August 2015
09:00-18:00
iGEM project
Digestion, ligation and transformation; PCR of pncB fusion; made more PSB1C3 backbone; Gel extraction; Nanodrop
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28th August 2015
11:00-18:00
iGEM project
Interlab study and safety form deadline; Redo ligation and transformation; PCR of PncB fusion (x2 on different settings); Gel of pncB
Week 11
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1st September 2015
iGEM Project
Redo Digestion, ligation and transformation
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2nd September 2015
09:00-11:00
iGEM project
Make plates, restreak plates, culture remaining cells on plates, G Blocks arrive. Ligation and Digestion
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3rd September 2015
11:00-18:00
iGEM project
Transformation and Figure out contaminants
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4th September 2015
09:00-18:00
iGEM project
Digestion, Ligation, Transformation
Week 12
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7th September 2015
iGEM project
Digestion and Ligation
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8th September 2015
09:00-18:00
iGEM project
Transformation
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9th September 2015
09:00-18:00
No labs concentration on presentation and powerpoint for Giant Jamboree
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10th September 2015
09:00-18:00
iGEM project
Re-streaks had unfortunately failed such that digestion and ligation were done again
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11th September 2015
09:00-18:00
iGEM project
Re-streaks of re try of digestions and ligations