Latest revision as of 01:28, 19 September 2015

Top

List of Note Books

Hijack Module (Fast robust oscillator) Notebook

Detection Chromoproteins Notebook

Detection Carotenoids Notebook (pdf)

Detection Enzyme Substrate Notebook

Termination notebook(pdf)

Hijack Module (Fast robust oscillator) Notebook

25^th June 2015

DH5α competent cells were transformed with the following biobricks:

BioBrick	Part	Backbone	Kit plate, Well
BBa_C0080	araC protein coding sequence	pSB1C3	3,11B
BBa_C0012	LacI protein coding sequence	pSB1C3	pSB1C3

The plates were kept at 37°C for 14 hours.

26^th June 2015

Transformation results:

Biobrick	Number of colonies
BBa_C0080	0
BBa_C0012	3

As number of colonies obtained was very low, the transformation of these biobricks was repeated.

27^th June 2015

Transformation results:

Biobrick	Number of colonies
BBa_C0080	3
BBa_C0012	60

Inoculation:

A colony from BBa_C0080 plate was inoculated in 7ml LB media + 7μL Chloramphenicol. Same was done for BBa_C0012. These cultures were kept at 37°C for 12 hours.

28^th June 2015

No growth was observed in both the liquid cultures.

A new colony was inoculated again.

29^th June 2015

Growth observed in the cultures of BBa_C0080 and BBa_C0012. Gylcerol stocks were made for both.

Plasmid isolation was done using Alkaline Lysis protocol for the 2 cultures. (1.5ml culture x 3)

Concentrations obtained from nanodrop:

Sample	Concentration (ng/μL)	A260/A280
BBa_C0080 (1)	3301.8	2.02
BBa_C0080 (2)	3670.9	1.98
BBa_C0080 (3)	2777.6	1.95
BBa_C0012 (1)	4560.0	1.97
BBa_C0012 (2)	4542.1	1.94
BBa_C0012 (3)	5123.5	1.61

A single digest was set for one of each of these plasmids:

(all volumes in μL)	BBa_C0012	BBa_C0080
PstI	0.3	0.3
10x NEBuffer 3	1	1
100x BSA	0.1	0.1
DNA	0.5	0.5
MilliQ H₂0	8.1	8.1
Total Volume	10	10

- Kept at 37°C for 2 hours

- Heat Inactivation: 80°C for 20 minutes

Gel Electrophoresis:

- Digested samples run on a 1% gel.

- Voltage: 100V

- Time run: 50 minutes

Both samples showed an RNA smear. This occurred as the Alkaline Lysis solution I did not have RNAse. After observing this smear, RNAse was added to the solution.

Re-inoculation of the two cultures was done

1^st July 2015

A plasmid isolation using Alkaline lysis was done.

Sample	Concentration (ng/μL)	A260/A280
BBa_C0080 (1)	4548.1	1.98
BBa_C0080 (2)	3411.9	2.01
BBa_C0080 (3)	3492.6	1.98
BBa_C0012 (1)	2258.4	1.99
BBa_C0012 (2)	2442.6	2.00
BBa_C0012 (3)	2691.4	1.97

A digest was set up with a sample of each plasmid and then run on a gel.

The gel showed bands at correct length, but the band intensity was extremely low and did not reflect the concentration obtained on nanodrop. A new method for plasmid isolation was needed.

2^nd July 2015

BBa_K094120 arrived from iGEM Headquarters and was inoculated in 5ml LB media + 5μL Ampicillin. Kept at 37°C for 12 hours.

3^rd July 2015

The liquid culture of BBa_K094120 was plated and kept overnight at 37°C

5^th July 2015

A single colony was inoculated for BBa_K094120. Kept at 37°C for 12 hours

6^th July 2015

Plasmid isolation of BBa_K094120 using Alkaline Lysis:

Sample	Concentration (ng/μL)	A260/A280
BBa_K094120 (1)	1050.8	1.88
BBa_K094120(2)	1493.7	1.95
BBa_K094120 (3)	1365.2	1.97

29^th July 2015

1% gel run with all samples for the 3 plasmids BBa_C0080, BBa_C0012, BBa_K094120.

All bands seen had low intensity which did not correlate to the concentration on nanodrop.

11^th August 2015

Revive glycerol stocks for BBa_C0080, BBa_K094120 and BBa_C0012. A 50 ml culture was inoculated for BBa_C0080 and BBa_C0012 for a Midiprep using Qiagen kit.

Kept at 37°C overnight

12^th August 2015

Midiprep using Qiagen Kit results for BBa_C0080 and BBa_C0012:

Sample	Concentration (ng/μL)	A260/A280
BBa_C0012	7.0	3.62
BBa_C0080	27.8	2.07

Since results obtained were very low, a new “Qiagen spin miniprep” kit was obtained.

17^th August 2015

Primary cultures used to make an overnight 5ml culture of BBa_C0080, BBa_C0012 and BBa_K094120.

18^th August 2015

Qiagen Spin Miniprep protocol followed. Results:

Sample	Concentration (ng/μL)	A260/A280
BBa_K094120	250.7	1.91
BBa_C0080	165.5	1.92
BBa_C0012	195.4	1.92

A digestion was set to check plasmid length:

(all volumes in μL)	BBa_C0080	BBa_C0012	BBa_K094120
DNA	2	2	2
EcoRI	0.2	0.2	0.2
10x EcoRI Buffer	1	1	1
100x BSA	0.1	0.1	0.1
MilliQ H₂0	6.4	6.4	6.4
Total Volume	10	10	10

Gel: 1% gel run at 100V for 40 minutes.

The bands obtained for all 3 plasmids were at the correct length. The uncut plasmid ran faster than cut plasmid.

21^st August 2015

Inoculated BBa_I13504 (GFP, used as reporter in oscillator construct) from previously transformed plate. Kept at 37°C for 12 hours

22^nd August 2015

- Miniprep of BBa_I13504:

Sample	Concentration (ng/μL)	A260/A280
BBa_I13504	122.4	1.94

- Transformation of the biobrick BBa_B0034 (RBS) in DH5α competent cells (Ampicillin antibiotic)

- Plates kept at 37°C for 14 hours

23^rd August 2015

Inoculated a colony for BBa_B0034 and kept at 37°C for 12 hours.

24^th August 2015

Plasmid Isolation using Qiagen Miniprep kit:

Sample	Concentration (ng/μL)	A260/A280
BBa_B0034	95.2	1.95

25^th August 2015

- Transformation of the biobrick BBa_B0015 (double terminator) in DH5α competent cells (Chloramphenicol antibiotic)

- Plates kept at 37°C for 14 hours

26^th August 2015

Transformation Result:

Number of colonies for BBa_B0015 = 8

- Inoculated BBa_B0015 in 5ml LB media + 5 μL Chloramphenicol

- Kept at 37°C for 12 hours

28^th August 2015

Plasmid Isolation of BBa_B0015 using Qiagen Miniprep kit:

Sample	Concentration (ng/μL)	A260/A280
BBa_B0034	72.3	1.95

29^th August 2015

Single Digest to check plasmid length:

(all volumes in μL)	BBa_B0015	BBa_I13504	BBa_B0034
DNA	4	2	2
EcoRI	0.2	0.2	0.2
10x EcoRI Buffer	1	1	1
100x BSA	0.1	0.1	0.1
MilliQ H₂0	5.7	5.7	5.7
Total Volume	10	10	10

Gel: 1% gel run with uncut and cut samples at 100V for 40 minutes.

The 3 single cut bands were at the right length.

6^th September 2015

Digestion for 3A assembly of constructs of the oscillator construct:

(all volumes given are in μL)	BBa_ K094120	BBa_ C0080	BBa_ C0012	BBa_ I13504	BBa_ B0034	BBa_ B0015	Ampicillin Backbone	Chloramphenicol Backbone	Kanamycin Backbone
DNA	3	3	3	3	4	5	10	10	10
10x NEBuffer 2	2.5	2.5	2.5	2.5	2.5	2.5	2.5	2.5	2.5
BSA	0.5	0.5	0.5	0.5	0.5	0.5	0.5	0.5	0.5
EcoRI	0.5	0.5	0.5	-	-	-	0.5	0.5	0.5
XbaI	-	-	-	0.5	0.5	0.5	-	-	-
SpeI	0.5	0.5	0.5	-	-	-	-	-	-
PstI	-	-	-	0.5	0.5	0.5	0.5	0.5	0.5
H₂0	13	13	13	13	12	11	6	6	6
Total	20	20	20	20	20	20	20	20	20

- Kept at 37°C for 5 hours

- Heat Inactivation at 80°C for 20 minutes

- The following devices were to be assembled:

- BBa_K094120 + BBa_B0034 + Chloramphenicol backbone = Device 2a

- BBa_C0080 + BBa_B0015 + Kanamycin backbone = Device 2b

- BBa_C0012 + + BBa_B0015 + Ampicillin backbone = Device 2c

- BBa_K094120 + BBa_I13504 + Kanamycin backbone = Device 2d

Ligation:

(all volumes in μL)	Device 2a	Device 2b	Device 2c	Device 2d
Ampicillin Backbone	-	-	2	-
Chloramphenicol Backbone	2	-	-	-
Kanamycin Backbone	-	2	-	2
BBa_K094120	2	-	-	2
BBa_B0034	2	-	-	-
BBa_C0080	-	2	-	-
BBa_B0015	-	2	2	-
BBa_C0012	-	-	2	-
BBa_I13504	-	-	-	2
T4 DNA ligase	0.5	0.5	0.5	0.5
Ligase Buffer	1	1	1	1
MilliQ H₂0	2.5	2.5	2.5	2.5
Total Volume	20	20	20	20

Ligation kept overnight at room temperature.

7^th September 2015

- Transformation of the ligation mix into DH5α cells

- Kept at 37°C overnight

8^th September 2015

Transformation results:

Ampicillin Negative Control : Lawn growth

Kanamycin Negative Control : Zero colonies

Plate	Number of Colonies
2a	10
2b	12
2c	Lawn growth
2d	5

Ampicillin degraded. 2c re-transformed

Inoculation of a single colony from each plate in 5ml LB media + 5μL antibiotic

Kept at 37°C for 12 hours

9^th September 2015

Miniprep by Qiagen Kit:

Sample	Concentration (ng/μL)	A260/A280
2a	283.4	1.91
2b	122.3	2.09
2d	226.6	1.96

Single Digest with EcoRI to check plasmid length:

Backbone-Backbone ligation was observed on the gel.

A new ligation was setup. This time insert:vector ratio was increased. Kept at 37°C for 2 hours.

These ligation mix were transformed into Dh5α competent cells.

10^th September 2015

Growth observed in negative control plates.

An overnight digestion was set for the individual plasmids again. Kept at 37°C for 12 hours

11^th September 2015

Ligation set for 2 hours and then transformed.

12^th September 2015

Colonies of device 2a, 2b and 2c obtained. No colonies on 2d.

3 colonies from each plate were inoculated, incubated for 12 hours at 37°C and miniprepped.

These were then digested with EcoRI to check the length of the plasmid on a gel.

The bands were as expected.

An overnight digest followed by 2 hour ligation was set to assemble to following device:

Device	Part1	Part2	Backbone
5a	2a	2b	pSB1A3
5b	2a	2c	pSB1K3

Two samples of 5b were assembled – one low copy and second high copy backbone

BBa_I13504 in Ampicillin backbone was chosen to assemble device 2d. This biobrick was digested and ligated.

Devices 5a, 5b and 2d were transformed.

13^th Spetember 2015

Transformation Results:
Negative Control: Zero

Plate	Number of Colonies
5a	15
5b (low copy)	70
5b (high copy)	26
2d	10

Three colonies were inoculated from each plate, incubated at 37°C and then miniprepped.

These were then single digested using EcorI to check on gel.

2d, 5a and 5b high copy showed correct bands.

18^th September 2015

2d construct was characterized by induction with different concentrations of IPTG and arabinose.

[top]

Chromoproteins

8th June, 2015

● The following biobrick parts were revived from the iGEM 2015 distribution plates using the iGEM protocol Standard protocol.

● Transformation was carried with Ultra-competent cells for DH5α strain of E.coli (with 200µL of LB).

● 200µL of the revived product was plated on LA+antibiotic.

● The plates were kept at 37℃ overnight (12-16 hours).

Part Number	Location in Kit	Backbone
J23110 (Constitutive Promoter)	Plate 4, Well 19D	Ampicillin
K592025 (RBS+amilCP)	Plate 1, Well 19C	Chloramphenicol
J04450 (RFP)	Plate 4, Well 2H	Ampicillin

Negative Control: For both antibiotics (Amp and Cam)

Positive Control: RFP

9th June, 2015

Part Number	Number of colonies
J23110 (Promoter)	Many colonies
K592025 (RBS+amilCP)	Many single colonies
J04450 (RFP)	Lawn growth
Negative Control	No colonies

● Next, the colonies were inoculated in 5mL LB+ 0.1% antibiotic.

● They were kept in an incubator at 37℃ overnight (12-16 hours) at around 180-200 rpm.

10th June, 2015

● Growth was observed in K592025, but no growth in J23110.

● Again a colony was picked from the plate with J23110 and inoculated in 5mL LB+ 0.1% antibiotic.

● They were kept in an incubator at 37℃ overnight (12-16 hours) at around 180-200 rpm.

11th June, 2015

● Again no growth was observed in J23110 (even after inoculating twice).

● Transformation was done again for J23110

● Different promoter was also transformed.

Part Number	Location in Kit	Backbone
J23119 (Constitutive Promoter)	Plate 3, Well 17O	Chloramphenicol

13th June, 2015

● The two promoters were tranformed with 200µL SOC media which was prepared using the iGEM protocol (earlier transformations with LB).

● 200µL of the revived product was plated on LA+antibiotic.

● The plates were kept at 37℃ overnight (12-16 hours).

14th June, 2015

Part Number	Number of colonies
J23110 (Promoter 1)	30
K592025 (RBS+amilCP)	3
J23119 (Promoter 2)	47
Negative Control	No colonies

● The colonies obtained were inoculated in 5mL LB+ 0.1% antibiotic.

● They were kept in an incubator at 37℃ overnight (12-16 hours) at around 180-200 rpm.

18th June, 2015

Plasmid isolation was carried out using the Alkaline Lysis Protocol from Sambrook and Maniatis.

Resuspended in TE Buffer.

Results of Nanodrop:

Sample	Vial Number	A260/A280	Concentration (ng/µL)
K592025	1	1.99	6691.9
	2	2.11	4749.5
	3	2.12	4158.3
J23119	1	2.10	4865.3
	2	2.09	2757.4
	3	2.10	4902.3
J23110	1	2.09	5121.8
	2	2.15	8148.0
	3	2.13	3223.8

● The alkaline lysis buffer I wasn’t added with RNase.

● The plasmid isolation was repeated with RNase added to Buffer I.

27th June, 2015

● The colonies were again inoculated in 5mL LB+ 0.1% antibiotic.

● They were kept in an incubator at 37℃ overnight (12-16 hours) at around 180-200 rpm.

28th June, 2015

● RNase (10mg/mL stock) was added to Alkaline Lysis Buffer I.

● Resuspension in milliQ water.

Results of Nanodrop:

Sample	Vial Number	A260/A280	Concentration (ng/µL)
K592025	1	2.13	56.9
	2	1.39	67.4
	3	2.06	58.5
J23119	1	2.16	34.1
	2	2.15	32.3
	3	1.39	116.1
J23110	1	2.08	158.4
	2	2.02	3222.1
	3	1.95	580.4

29th June, 2015

● The colonies were again inoculated in 5mL LB+ 0.1% antibiotic.

● They were kept in an incubator at 37℃ overnight (12-16 hours) at around 180-200 rpm.

30th June-10th July, 2015

Again, alkaline lysis was done.

Results of Nanodrop:

Date	Sample	Vial Number	A260/A280	Concentration (ng/µL)
30/6	J23119	1	1.94	2376.8
30/6	J23119	2	1.95	2301.2
30/6	J23119	3	1.88	1662.9
8/7	J23119	1	1.99	1616.5
8/7	J23110	2	1.98	618.9
10/7	K592025	1	1.96	1641.1
10/7	K592025	2	1.97	1457.0
10/7	K592025	3	1.99	1404.6

26th August and 3rd September

Results for Nanodrop by Qiagen spin mini kit

Sample	A260/A280	Concentration (ng/μL)
J23119	2.04	121.1
J23119	1.89	329.3
K592029	1.96	158
K592029	1.88	407.3

26th August

Single digestion reaction

(in μL)	K592025	J23119
EcoRI	0.3	0.3
DNA	2	2
10XEcoRI Buffer	1	1
100X BSA	0.1	0.1
Distilled water	6.6	6.6
Total Volume	10	10

27th August

3A assembly double digestion

(in μL)	Promoter(J23119)	amilCP(K592025)	Linear backbone
DNA	2	2	10
10X Buffer2	2.5	2.5	2.5
100X BSA	0.5	0.5	0.5
EcoRI	0.5	-	0.5
SpeI	0.5	-	-
PstI	-	0.5	0.5
XbaI	-	0.5	-
DpnI	-	-	0.5
Distlled water	14	14	5.5
Total Volume	20	20	20

Kept at 37°C for 4 hours, heat-kill at 80°C for 20min.

Colonies obtained (~10), were inoculated and miniprep was done with qiagen kit.

Table3: Double digestion for 2A assembly

2A assembly Double digestion:

(in μL)	Promoter(J23119)	amilCP(K592025)
DNA	7.8	7
10X Buffer2	1	1
100X BSA	0.2	0.2
SpeI	0.5	0.5
PstI	0.5	-
XbaI	-	0.5
Distilled water	0	0.8
Total Volume	10	10

[top]

Detection Enzyme Substrate Notebook

DETECTION: Enzyme substrate reaction

Parts:

	Biobrick	Part	Size(bp)	Location
1	BBa_K1509003	Constitutive promoter	2129	Ordered from iGEM HQ
2	BBa_K823016	RBS + full length LacZ	5163	Kit plate 2

5/08/15

Transformation of BBa_K823016 (LacZ gene along RBS) plasmid in DH5α competent cells

Protocol: iGEM Transformation protocol

Master plate is made

6/08/15

Observed colonies on the plate

12/08/15

Liquid culture is prepared for BBa_K823016

Protocol

5ml of LB in 15ml falcon
Add 5µl of cam
Pick a colony from the plate and dip in falcon
Incubate for 12-14hrs

22/08/15

BBa_K1509003 (promoter) is ordered and got from iGEM HQ. NEB10 is the cell line

Preparation of liquid culture

5ml LB in 15ml falcon
Add 5µl of cam
A loop of nichrome dipped in NEB10 cells containing promoter plasmid
Incubate for 12-14hrs

23/08/15

Streaking of plate

Prepare a cam media plate of 20ml LA
Streak the plate using previously prepared liquid culture
Overnight Incubation

24/08/15

Grow liquid stalk culture

5ml LB in 15ml falcon
Add 5µl of cam
Pick a colony from master plate and dip in media
Overnight incubation

Preparation of glycerol stalk

0.5ml 80% glycerol in cryotube
0.5ml bacterial culture
Flash freeze it using liquid nitrogen

24/08/15

Plasmid isolation by alkaline lysis

Chemicals required:

CAM antibiotic
LB media
Alkaline lysis buffer 1
Alkaline lysis buffer 2
Alkaline lysis buffer 3
Ethanol
Phenol: Chloroform (1:1)
STE
TE with RNase A

Protocol

Inoculate 5ml of rich media containing the appropriate antibiotic (CAM)
Incubate culture overnight at 37 deg
Pour 5ml of culture into a microfuge tube
Centrifuge in microfuge for 3 min at 4deg
Remove media by aspiration leaving bacterial pellet as dry as possible
Resuspend the pellet in 100microl of ice-cold alkaline lysis solution 1 by vortexing
Add 200microl of alkaline lysis solution 2 , mix with inverting
Add 150 microl ice-cold alkaline lysis solution 3 mix with invertexing
Store tube on ice for 3-5 min
Centrifuge bacterial lysate ,5 min 13000rpm at 4deg in microfuge
Transfer supernatant to fresh tube
Add equal volume of phenol: chloroform
Mix the organic and aqueous phase by vortexing
Centrifuge at maximum speed 2 min at 4deg
Transfer the aqueous upper layer to fresh tube
Add 2 volume of ethanol
Mix by vortexing for 2 min
Centrifuge for 5min at 4deg
Remove supernatant by aspiration
Remove all liquid keeping inverted position
Add 1ml of ethanol to pellet
Centrifuge for 2min at 4deg
Remove all supernatant by aspiration
Remove ethanol beads and evaporate it at room temperature
Dissolve nucleic acid in 50microl of TE containing RNases A
Vortex the solution gently for 2 sec
Store DNA solution at -20deg

Result: no DNA pellet after precipitation by ethanol

Comment: Most probably step 12 is done in incorrect way

Inoculation of a colony of BBa_K823016 and BBa_K1509003

25/08/15

Plasmid isolation of BBa_K1509003 using QIAGEN miniprep kit

Protocol: QIAGEN Miniprep gravity column is used

Result: No DNA after precipitation by isopropanol addition

26/08/15

Inoculation of BBa_K823016 and BBa_K1509003

Plasmid isolation using QIAGEN Miniprep spin column

Protocol: QIAGEN Miniprep spin column protocol is used

Results:

part	Concentration(ng/µl)
BBa_K823016	52.2
BBa_K1509003	38.7

Inoculation of BBa_K823016 and BBa_K1509003 for alkaline lysis

27/08/15

Plasmid isolation using Alkaline lysis Miniprep in AB Lab

Chemicals Required: Alkaline lysis buffer 1, 2, 3 buffer 2 freshly prepared, RNases

Protocol: standard protocol is used from sambrook molecular cloning book as mentioned earlier

Result

part	Concentration(ng/µl)
BBa_K823016	2650.8
BBa_K1509003	4080.3

This has high RNA contamination

Gel Electrophoresis is done which shows high contamination of RNA

2/09/15

Inoculation of both BBa_K823016 and BBa_K1509003

Plasmid isolation using QIAGEN Miniprep spin column

Protocol: QIAGEN Miniprep spin column protocol is used

Result:

part	Concentration(ng/µl)
BBa_K823016	275.2
BBa_K1509003	87.3

Gel Electrophoresis is done to check isolated plasmid

8/09/15

Inoculation of BBa_K1509003 in 5ml LB and 5µl cam

9/09/15

Gel Electrophoresis (1%)

Chemicals required: 0.5gm agarose, 50ml TAE, 5µl EtBr

Inoculation of both BBa_K823016 and BBa_K1509003 in 5ml LB each

10/09/15

Plasmid isolation using QIAGEN Miniprep spin column

Protocol: QIAGEN Miniprep spin column protocol is used

Result:

part	Concentration(ng/µl)
BBa_K823016	735
BBa_K1509003	234

Single cut digestion

BBa_K823016: 1µl

BBa_K1509003:3µl

	BBa_K823016 (µl)	BBa_K1509003 (µl)
DNA	1	3
ECoR1 buffer	1	1
ECoR1 enzyme	0.5	0.5
dH2O	7.5	5.5
Total	10	10

Gel Electrophoresis: Fig1

12/09/15

Fig2

Confirmation of plasmid isolated

12/09/15

Digestion for 3A assembly

pSB1K3 which is a Kan backbone used for 3A assembly

		BBa_K823016 (µl)	BBa_K1509003 (µl)
1	NEB buffer 2	5	5
2	BSA	0.5	0.5
3	EcoR1-HF	0.5	0.5
4	Spel	0.5	0.5
5	dH20	18.5	18.5
6	Total	20	20

Protocol: standard iGEM 3A assembly protocol is used

13/09/15

Ligation using 3A assembly of biobricks

Protocol: standard iGEM 3A assembly protocol is used

15/09/15

Transformation of ligated parts

Protocol: IGEM transformation protocol is used

Result: No colony is observed on both KAN media plates

Comment: most probably failure of digestion

16/09/15

Inoculation of both BBa_K823016 and BBa_K1509003

Plasmid isolation using QIAGEN Miniprep spin column

Protocol: QIAGEN Miniprep spin column protocol is used

Result:

part	Concentration(ng/µl)
BBa_K823016	102.3
BBa_K1509003	65.5

[top]

@@ Line 9: / Line 9: @@
 <h2>List of Note Books</h2>
+<a name="top"></a>
 <ul>
-      <li> <a href="#Hijack_Module">Hijack Module (Fast robust oscillator) Notebook< /a> </li>
+      <a href="#Hijack_Module"><li> Hijack Module (Fast robust oscillator) Notebook</li></a>
-      <li><a>Hijack Module (Fast robust oscillator) Notebook< /a> </li>
+      <a href="#Chromoproteins"><li>Detection Chromoproteins Notebook </li>  </a>
+     <a href="https://static.igem.org/mediawiki/2015/2/29/IISER_PUNE_Notebook_Carotenoids.pdf"><li>Detection Carotenoids Notebook (pdf)</li>  </a>
+     <a href="#Enzyme_Substrate"><li> Detection Enzyme Substrate Notebook</li></a>
+     <a href="https://static.igem.org/mediawiki/2015/9/97/IISER_PUNE_Notebook_Terminator.pdf"><li>Termination notebook(pdf)</li> </a>
@@ Line 3,526: / Line 3,535: @@
 </div>
+[<a href="#top">top</a>]
+<div class=WordSection1>
+<a name="Chromoproteins"></a>
+<p> <h1> Chromoproteins</h1></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>8th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l3 level1 lfo7'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The following <span class=SpellE>biobrick</span> parts
+were revived from the <span class=SpellE><b style='mso-bidi-font-weight:normal'>iGEM</b></span><b
+style='mso-bidi-font-weight:normal'> 2015 distribution plates </b>using the <span
+class=SpellE>iGEM</span> protocol Standard protocol.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l3 level1 lfo7'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Transformation was carried with Ultra-competent cells
+for <b style='mso-bidi-font-weight:normal'>DH5&#945; </b>strain of <b
+style='mso-bidi-font-weight:normal'><i style='mso-bidi-font-style:normal'>E.coli
+</i></b>(with 200µL of LB). </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l3 level1 lfo7'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>200µL of the revived product was plated on <span
+class=SpellE>LA+antibiotic</span>.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l3 level1 lfo7'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The plates were kept at 37&#8451; overnight (12-16
+hours).</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a border=1 cellspacing=0 cellpadding=0 width=624 style='border-collapse:
+ collapse;mso-table-layout-alt:fixed;border:none;mso-border-alt:solid black 1.0pt;
+ mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
+.0pt solid black;mso-border-insidev:1.0pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Part Number</b></p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Location in Kit</b></p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Backbone</b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><span
+  style='font-size:9.5pt;background:white;mso-highlight:white'>J23110
+  (Constitutive Promoter)</span></p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Plate 4,
+  Well 19D</p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Ampicillin</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025 (<span
+  class=SpellE>RBS+amilCP</span>)</p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Plate 1,
+  Well 19C</p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Chloramphenicol</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;mso-yfti-lastrow:yes'>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J04450
+  (RFP)</p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Plate 4,
+  Well 2H</p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Ampicillin</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>Negative Control</b>:
+For both antibiotics (Amp and Cam)</p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>Positive Control</b>:
+RFP</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>9th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a0 border=1 cellspacing=0 cellpadding=0 width=624
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black 1.0pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:1.0pt solid black;mso-border-insidev:1.0pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Part Number</b></p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Number of colonies</b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><span
+  style='font-size:9.5pt;background:white;mso-highlight:white'>J23110
+  (Promoter)</span></p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Many
+  colonies</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025 (<span
+  class=SpellE>RBS+amilCP</span>)</p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Many single
+  colonies</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J04450
+  (RFP)</p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Lawn growth</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Negative
+  Control</p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>No colonies</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Next, the colonies were inoculated in 5mL LB+ 0.1%
+antibiotic.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>They were kept in an incubator at 37&#8451; overnight
+(12-16 hours) at around 180-200 rpm.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>10th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l0 level1 lfo2'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Growth was observed in K592025, but no growth in
+J23110.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l0 level1 lfo2'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Again a colony was picked from the plate with J23110
+and inoculated in 5mL LB+ 0.1% antibiotic.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l0 level1 lfo2'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>They were kept in an incubator at 37&#8451; overnight
+(12-16 hours) at around 180-200 rpm. </p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>11th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l2 level1 lfo3'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Again no growth was observed in J23110 (even after
+inoculating twice).</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l2 level1 lfo3'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Transformation was done again for J23110</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l2 level1 lfo3'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Different promoter was also transformed.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a1 border=1 cellspacing=0 cellpadding=0 width=624
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black 1.0pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:1.0pt solid black;mso-border-insidev:1.0pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Part Number</b></p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Location in Kit</b></p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Backbone</b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
+  <td width=208 valign=top style='width:156.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><span
+  style='font-size:9.5pt;background:white;mso-highlight:white'>J23119
+  (Constitutive Promoter)</span></p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Plate 3,
+  Well 17O</p>
+  </td>
+  <td width=208 valign=top style='width:156.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Chloramphenicol</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>13th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l1 level1 lfo4'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The two promoters were <span class=SpellE>tranformed</span>
+with 200µL SOC media which was prepared using the <span class=SpellE>iGEM</span>
+protocol (earlier transformations with LB).</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l1 level1 lfo4'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>200µL of the revived product was plated on <span
+class=SpellE>LA+antibiotic</span>.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l1 level1 lfo4'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The plates were kept at 37&#8451; overnight (12-16
+hours).</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>14th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a2 border=1 cellspacing=0 cellpadding=0 width=624
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black 1.0pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:1.0pt solid black;mso-border-insidev:1.0pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Part Number</b></p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Number of colonies</b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><span
+  style='font-size:9.5pt;background:white;mso-highlight:white'>J23110 (Promoter
+)</span></p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>30</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025 (<span
+  class=SpellE>RBS+amilCP</span>)</p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23119
+  (Promoter 2)</p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>47</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes'>
+  <td width=312 valign=top style='width:234.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>Negative
+  Control</p>
+  </td>
+  <td width=312 valign=top style='width:234.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>No colonies</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The colonies obtained were inoculated in 5mL LB+ 0.1%
+antibiotic.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>They were kept in an incubator at 37&#8451; overnight
+(12-16 hours) at around 180-200 rpm.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>18th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal>Plasmid isolation was carried out using the <b
+style='mso-bidi-font-weight:normal'>Alkaline Lysis Protocol </b>from <span
+class=SpellE>Sambrook</span> and <span class=SpellE>Maniatis</span>.</p>
+<p class=MsoNormal><span class=SpellE><span class=GramE>Resuspended</span></span><span
+class=GramE> in TE Buffer.</span></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>Results of <span
+class=SpellE>Nanodrop</span>:</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a3 border=1 cellspacing=0 cellpadding=0 width=624
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black 1.0pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:1.0pt solid black;mso-border-insidev:1.0pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:23.0pt'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt;height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Sample</b></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Vial Number</b></p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>A260/A280</b></p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Concentration (ng/µL)</b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1.99</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>6691.9</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.11</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>4749.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.12</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>4158.3</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23119</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.10</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>4865.3</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.09</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2757.4</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.10</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>4902.3</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23110</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.09</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>5121.8</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.15</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>8148.0</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:9;mso-yfti-lastrow:yes'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.13</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3223.8</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l5 level1 lfo5'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The alkaline lysis buffer I wasn’t added with RNase. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l5 level1 lfo5'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The plasmid isolation was repeated with RNase added to
+Buffer I.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>27th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The colonies were again inoculated in 5mL LB+ 0.1%
+antibiotic.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>They were kept in an incubator at 37&#8451; overnight
+(12-16 hours) at around 180-200 rpm.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>28th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l4 level1 lfo6'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>RNase (10mg/mL stock) was added to Alkaline Lysis
+Buffer I.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l4 level1 lfo6'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Resuspension in <span class=SpellE>milliQ</span> water.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>Results of <span
+class=SpellE>Nanodrop</span>:</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a4 border=1 cellspacing=0 cellpadding=0 width=624
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black 1.0pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:1.0pt solid black;mso-border-insidev:1.0pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:23.0pt'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt;height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Sample</b></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Vial Number</b></p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>A260/A280</b></p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Concentration (ng/µL)</b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.13</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>56.9</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1.39</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>67.4</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.06</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>58.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23119</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.16</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>34.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.15</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>32.3</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1.39</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>116.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23110</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.08</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>158.4</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2.02</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3222.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:9;mso-yfti-lastrow:yes'>
+  <td width=156 valign=top style='width:117.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1.95</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>580.4</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>29th June, 2015</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>The colonies were again inoculated in 5mL LB+ 0.1%
+antibiotic.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:36.0pt;mso-add-space:auto;
+text-indent:-18.0pt;mso-list:l6 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>They were kept in an incubator at 37&#8451; overnight
+(12-16 hours) at around 180-200 rpm.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>30th June-10th July,
+</b></p>
+<p class=MsoNormal>Again, alkaline lysis was done.</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>Results of <span
+class=SpellE>Nanodrop</span>:</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a5 border=1 cellspacing=0 cellpadding=0 width=624
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black 1.0pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:1.0pt solid black;mso-border-insidev:1.0pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:23.0pt'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt;height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Date</b></p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Sample</b></p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Vial Number</b></p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>A260/A280</b></p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:23.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'><b
+  style='mso-bidi-font-weight:normal'>Concentration (ng/µL)</b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>30/6</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23119</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.94</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2376.8</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>30/6</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23119</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.95</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2301.2</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>30/6</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23119</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.88</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1662.9</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>8/7</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23119</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.99</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1616.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>8/7</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>J23110</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.98</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>618.9</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>10/7</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>1</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.96</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1641.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>10/7</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>2</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.97</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1457.0</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8;mso-yfti-lastrow:yes'>
+  <td width=78 valign=top style='width:58.5pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>10/7</p>
+  </td>
+  <td width=78 valign=top style='width:58.5pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>K592025</p>
+  </td>
+  <td width=124 valign=top style='width:93.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-pagination:none'>3</p>
+  </td>
+  <td width=160 valign=top style='width:120.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.99</p>
+  </td>
+  <td width=184 valign=top style='width:138.0pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black 1.0pt;mso-border-left-alt:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1404.6</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><span style='mso-no-proof:yes'><!--[if gte vml 1]><v:shapetype
+ id="_x0000_t75" coordsize="21600,21600" o:spt="75" o:preferrelative="t"
+ path="m@4@5l@4@11@9@11@9@5xe" filled="f" stroked="f">
+ <v:stroke joinstyle="miter"/>
+ <v:formulas>
+  <v:f eqn="if lineDrawn pixelLineWidth 0"/>
+  <v:f eqn="sum @0 1 0"/>
+  <v:f eqn="sum 0 0 @1"/>
+  <v:f eqn="prod @2 1 2"/>
+  <v:f eqn="prod @3 21600 pixelWidth"/>
+  <v:f eqn="prod @3 21600 pixelHeight"/>
+  <v:f eqn="sum @0 0 1"/>
+  <v:f eqn="prod @6 1 2"/>
+  <v:f eqn="prod @7 21600 pixelWidth"/>
+  <v:f eqn="sum @8 21600 0"/>
+  <v:f eqn="prod @7 21600 pixelHeight"/>
+  <v:f eqn="sum @10 21600 0"/>
+ </v:formulas>
+ <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>
+ <o:lock v:ext="edit" aspectratio="t"/>
+</v:shapetype><v:shape id="image04.png" o:spid="_x0000_i1027" type="#_x0000_t75"
+ style='width:453.75pt;height:288.75pt;visibility:visible;mso-wrap-style:square'>
+ <v:imagedata src="Chromoproteinsw_files/image001.png" o:title=""/>
+</v:shape><![endif]--><![if !vml]><img width=605 height=385
+src="Chromoproteinsw_files/image002.gif" v:shapes="image04.png"><![endif]></span></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a6 border=0 cellspacing=0 cellpadding=0 width=600
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;mso-yfti-tbllook:
+;mso-padding-alt:0cm 5.4pt 0cm 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:81.0pt'>
+  <td width=576 valign=top style='width:432.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:81.0pt'>
+  <p class=MsoNormal style='mso-pagination:none'><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=24 valign=top style='width:18.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt;
+  height:81.0pt'>
+  <p class=MsoNormal><o:p>&nbsp;</o:p></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
+  <td width=576 valign=top style='width:432.0pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal><o:p>&nbsp;</o:p></p>
+  </td>
+  <td width=24 valign=top style='width:18.0pt;border:solid black 1.0pt;
+  mso-border-alt:solid black .75pt;background:white;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal><o:p>&nbsp;</o:p></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>26th August and 3rd
+September</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal>Results for <span class=SpellE>Nanodrop</span> by <span
+class=SpellE>Qiagen</span> spin mini kit</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a7 border=1 cellspacing=0 cellpadding=0 width=592
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black .75pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:.75pt solid black;mso-border-insidev:.75pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=132 valign=top style='width:99.0pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Sample</p>
+  </td>
+  <td width=175 valign=top style='width:131.25pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>A260/A280</p>
+  </td>
+  <td width=285 valign=top style='width:213.75pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Concentration (ng/<span
+  class=SpellE>&#956;L</span>)</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=132 valign=top style='width:99.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>J23119</p>
+  </td>
+  <td width=175 valign=top style='width:131.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2.04</p>
+  </td>
+  <td width=285 valign=top style='width:213.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>121.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=132 valign=top style='width:99.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>J23119</p>
+  </td>
+  <td width=175 valign=top style='width:131.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.89</p>
+  </td>
+  <td width=285 valign=top style='width:213.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>329.3</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=132 valign=top style='width:99.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>K592029</p>
+  </td>
+  <td width=175 valign=top style='width:131.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.96</p>
+  </td>
+  <td width=285 valign=top style='width:213.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>158</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes'>
+  <td width=132 valign=top style='width:99.0pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>K592029</p>
+  </td>
+  <td width=175 valign=top style='width:131.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1.88</p>
+  </td>
+  <td width=285 valign=top style='width:213.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>407.3</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><span style='mso-no-proof:yes'><!--[if gte vml 1]><v:shape
+ id="image05.png" o:spid="_x0000_i1026" type="#_x0000_t75" style='width:468pt;
+ height:282pt;visibility:visible;mso-wrap-style:square'>
+ <v:imagedata src="Chromoproteinsw_files/image003.png" o:title=""/>
+</v:shape><![endif]--><![if !vml]><img width=624 height=376
+src="Chromoproteinsw_files/image004.gif" v:shapes="image05.png"><![endif]></span></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><span style='mso-no-proof:yes'><!--[if gte vml 1]><v:shape
+ id="image02.png" o:spid="_x0000_i1025" type="#_x0000_t75" style='width:468pt;
+ height:279pt;visibility:visible;mso-wrap-style:square'>
+ <v:imagedata src="Chromoproteinsw_files/image005.png" o:title=""/>
+</v:shape><![endif]--><![if !vml]><img width=624 height=372
+src="Chromoproteinsw_files/image006.gif" v:shapes="image02.png"><![endif]></span></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>26th August</b></p>
+<p class=MsoNormal>Single digestion reaction</p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<table class=a8 border=1 cellspacing=0 cellpadding=0 width=592
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black .75pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:.75pt solid black;mso-border-insidev:.75pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=199 valign=top style='width:149.25pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>(in <span class=SpellE>&#956;L</span>)</p>
+  </td>
+  <td width=198 valign=top style='width:148.5pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>K592025</p>
+  </td>
+  <td width=195 valign=top style='width:146.25pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>J23119</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=199 valign=top style='width:149.25pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>EcoRI</span></p>
+  </td>
+  <td width=198 valign=top style='width:148.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.3</p>
+  </td>
+  <td width=195 valign=top style='width:146.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.3</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=199 valign=top style='width:149.25pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>DNA</p>
+  </td>
+  <td width=198 valign=top style='width:148.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2</p>
+  </td>
+  <td width=195 valign=top style='width:146.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=199 valign=top style='width:149.25pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10XEcoRI Buffer</p>
+  </td>
+  <td width=198 valign=top style='width:148.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1</p>
+  </td>
+  <td width=195 valign=top style='width:146.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=199 valign=top style='width:149.25pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>100X BSA</p>
+  </td>
+  <td width=198 valign=top style='width:148.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.1</p>
+  </td>
+  <td width=195 valign=top style='width:146.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=199 valign=top style='width:149.25pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Distilled water</p>
+  </td>
+  <td width=198 valign=top style='width:148.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>6.6</p>
+  </td>
+  <td width=195 valign=top style='width:146.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>6.6</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6;mso-yfti-lastrow:yes'>
+  <td width=199 valign=top style='width:149.25pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Total Volume</p>
+  </td>
+  <td width=198 valign=top style='width:148.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10</p>
+  </td>
+  <td width=195 valign=top style='width:146.25pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><b style='mso-bidi-font-weight:normal'>27th August</b></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal>3A assembly double digestion</p>
+<table class=a9 border=1 cellspacing=0 cellpadding=0 width=624
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black .75pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:.75pt solid black;mso-border-insidev:.75pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>(in <span class=SpellE>&#956;L</span>)</p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Promoter(J23119)</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>amilCP</span>(K592025)</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Linear backbone</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>DNA</p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10X Buffer2</p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2.5</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2.5</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>2.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>100X BSA</p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>EcoRI</span></p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>SpeI</span></p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>PstI</span></p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>XbaI</span></p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>DpnI</span></p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:9'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>Distlled</span>
+  water</p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>14</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>14</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>5.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:10;mso-yfti-lastrow:yes'>
+  <td width=132 valign=top style='width:98.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Total Volume</p>
+  </td>
+  <td width=175 valign=top style='width:130.95pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>20</p>
+  </td>
+  <td width=167 valign=top style='width:125.2pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>20</p>
+  </td>
+  <td width=151 valign=top style='width:113.05pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>20</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal>Kept at 37°C for 4 hours<span class=GramE>,<span
+style='mso-spacerun:yes'>  </span>heat</span>-kill at 80°C for 20min.</p>
+<p class=MsoNormal>Colonies obtained (~10)<span class=GramE>,<span
+style='mso-spacerun:yes'>  </span>were</span> inoculated and <span
+class=SpellE>miniprep</span> was done with <span class=SpellE>qiagen</span>
+kit.</p>
+<p class=MsoNormal><span style='mso-spacerun:yes'> </span></p>
+<table class=aa border=0 cellspacing=0 cellpadding=0 width=606
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;mso-yfti-tbllook:
+;mso-padding-alt:0cm 5.4pt 0cm 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
+  <td width=606 valign=top style='width:454.75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Table3: Double digestion for 2A
+  assembly</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal>2A assembly Double digestion:</p>
+<p class=MsoNormal><span style='mso-spacerun:yes'> </span></p>
+<table class=ab border=1 cellspacing=0 cellpadding=0 width=592
+ style='border-collapse:collapse;mso-table-layout-alt:fixed;border:none;
+ mso-border-alt:solid black .75pt;mso-yfti-tbllook:1536;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
+ mso-border-insideh:.75pt solid black;mso-border-insidev:.75pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>(in <span class=SpellE>&#956;L</span>)</p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Promoter(J23119)</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border:solid black 1.0pt;
+  border-left:none;mso-border-left-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>amilCP</span>(K592025)</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>DNA</p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>7.8</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>7</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10X Buffer2</p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>100X BSA</p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.2</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.2</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>SpeI</span></p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>PstI</span></p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>XbaI</span></p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>-</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.5</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Distilled water</p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>0.8</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8;mso-yfti-lastrow:yes'>
+  <td width=177 valign=top style='width:132.75pt;border:solid black 1.0pt;
+  border-top:none;mso-border-top-alt:solid black .75pt;padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>Total Volume</p>
+  </td>
+  <td width=209 valign=top style='width:156.75pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10</p>
+  </td>
+  <td width=206 valign=top style='width:154.5pt;border-top:none;border-left:
+  none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .75pt;mso-border-left-alt:solid black .75pt;
+  padding:5.0pt 5.0pt 5.0pt 5.0pt'>
+  <p class=MsoNormal style='line-height:normal'>10</p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span style='mso-spacerun:yes'> </span></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
+</div>
+[<a href="top">top</a>]
+<div id="EnzymeSubstrate">
+<a name="Enzyme_Substrate"></a>
+<p> <h1> Detection Enzyme Substrate Notebook</h1></p>
+<p class="c0 c7"><a name="h.gjdgxs"></a></p><p class="c0"><span class="c28 c10">DETECTION: Enzyme substrate reaction</span></p><p class="c0"><span class="c10 c28">Parts: </span></p><a href="#" name="7258759fbe131b2bd7e57ada4ee7574bf50e6c65"></a><a href="#" name="0"></a><table cellpadding="0" cellspacing="0" class="c18"><tbody><tr class="c27"><td class="c21" colspan="1" rowspan="1"><p class="c1 c0 c7"><span class="c5 c23 c10"></span></p></td><td class="c29" colspan="1" rowspan="1"><p class="c1 c0"><span class="c10 c25">Biobrick </span></p></td><td class="c24" colspan="1" rowspan="1"><p class="c0 c1"><span class="c5 c10 c26">Part</span></p></td><td class="c30" colspan="1" rowspan="1"><p class="c1 c0"><span class="c25 c10">Size(bp) </span></p></td><td class="c16" colspan="1" rowspan="1"><p class="c1 c0"><span class="c25 c10">Location </span></p></td></tr><tr class="c7"><td class="c21" colspan="1" rowspan="1"><p class="c1 c0"><span class="c25 c10">1 </span></p></td><td class="c29" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c23 c10">BBa_K1509003 </span></p></td><td class="c24" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c10 c23">Constitutive promoter</span></p></td><td class="c30" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c23 c10">2129 </span></p></td><td class="c16" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c23 c10">Ordered from iGEM HQ </span></p></td></tr><tr class="c27"><td class="c21" colspan="1" rowspan="1"><p class="c1 c0"><span class="c25 c10">2 </span></p></td><td class="c29" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c23 c10">BBa_K823016 </span></p></td><td class="c24" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c23 c10">RBS + full length LacZ</span></p></td><td class="c30" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c23 c10">5163 </span></p></td><td class="c16" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c23 c10">Kit plate 2 </span></p></td></tr></tbody></table><p class="c0 c7"><span class="c10"></span></p><p class="c0"><span>5/08/15</span></p><p class="c0"><span>Transformation of BBa_K823016 (LacZ gene along RBS) plasmid in DH5&alpha; competent cells</span></p><p class="c0"><span>Protocol: iGEM Transformation protocol</span></p><p class="c0"><span>Master plate is made</span></p><p class="c0"><span>6/08/15</span></p><p class="c0"><span>Observed colonies on the plate</span></p><p class="c0"><span>12/08/15</span></p><p class="c0"><span>Liquid culture is prepared for BBa_K823016</span></p><p class="c0"><span>Protocol</span></p><ul class="c19 lst-kix_list_1-0 start"><li class="c4 c0"><span class="c3">5ml of LB in 15ml falcon</span></li><li class="c4 c0"><span class="c3">Add 5&micro;l of cam</span></li><li class="c4 c0"><span class="c3">Pick a colony from the plate and dip in falcon</span></li><li class="c4 c0"><span class="c3">Incubate for 12-14hrs</span></li></ul><p class="c0"><span>22/08/15</span></p><p class="c0"><span>BBa_K1509003 (promoter) is ordered and got from iGEM HQ. NEB10 is the cell line</span></p><p class="c0"><span>Preparation of liquid culture</span></p><ul class="c19 lst-kix_list_2-0 start"><li class="c4 c0"><span class="c3">5ml LB in 15ml falcon</span></li><li class="c4 c0"><span class="c3">Add 5&micro;l of cam </span></li><li class="c4 c0"><span class="c3">A loop of nichrome dipped in NEB10 cells containing promoter plasmid</span></li><li class="c4 c0"><span class="c3">Incubate for 12-14hrs</span></li></ul><p class="c0 c7"><span></span></p><p class="c0"><span>23/08/15</span></p><p class="c0"><span>Streaking of plate</span></p><ul class="c19 lst-kix_list_3-0 start"><li class="c4 c0"><span class="c3">Prepare a cam media plate of 20ml LA </span></li><li class="c4 c0"><span class="c3">Streak the plate using previously prepared liquid culture</span></li><li class="c4 c0"><span class="c3">Overnight Incubation </span></li></ul><p class="c0 c7"><span></span></p><p class="c0"><span>24/08/15</span></p><p class="c0"><span>Grow liquid stalk culture</span></p><ul class="c19 lst-kix_list_4-0 start"><li class="c4 c0"><span class="c3">5ml LB in 15ml falcon</span></li><li class="c4 c0"><span class="c3">Add 5&micro;l of cam</span></li><li class="c4 c0"><span class="c3">Pick a colony from master plate and dip in media</span></li><li class="c4 c0"><span class="c3">Overnight incubation</span></li></ul><p class="c0"><span>Preparation of glycerol stalk</span></p><ul class="c19 lst-kix_list_5-0 start"><li class="c4 c0"><span class="c3">0.5ml 80% glycerol in cryotube</span></li><li class="c4 c0"><span class="c3">0.5ml bacterial culture</span></li><li class="c4 c0"><span class="c3">Flash freeze it using liquid nitrogen</span></li></ul><p class="c0"><span>24/08/15</span></p><p class="c0"><span>Plasmid isolation by alkaline lysis</span></p><p class="c0"><span>Chemicals required:</span></p><ol class="c19 lst-kix_list_6-0 start" start="1"><li class="c4 c0"><span class="c3">CAM antibiotic</span></li><li class="c4 c0"><span class="c3">LB media</span></li><li class="c4 c0"><span class="c3">Alkaline lysis buffer 1</span></li><li class="c4 c0"><span class="c3">Alkaline lysis buffer 2</span></li><li class="c4 c0"><span class="c3">Alkaline lysis buffer 3</span></li><li class="c4 c0"><span class="c3">Ethanol</span></li><li class="c4 c0"><span class="c3">Phenol: Chloroform (1:1)</span></li><li class="c4 c0"><span class="c3">STE</span></li><li class="c4 c0"><span class="c3">TE with RNase A</span></li></ol><p class="c0"><span>Protocol</span></p><ol class="c19 lst-kix_list_7-0 start" start="1"><li class="c4 c0"><span class="c3">Inoculate 5ml of rich media containing the appropriate antibiotic (CAM)</span></li><li class="c4 c0"><span class="c3">Incubate culture overnight at 37 deg</span></li><li class="c4 c0"><span class="c3">Pour 5ml of culture into a microfuge tube</span></li><li class="c4 c0"><span class="c3">Centrifuge in microfuge for 3 min at 4deg </span></li><li class="c4 c0"><span class="c3">Remove media by aspiration leaving bacterial pellet as dry as possible</span></li><li class="c4 c0"><span class="c3">Resuspend the pellet in 100microl of ice-cold alkaline lysis solution 1 by vortexing </span></li><li class="c4 c0"><span class="c3">Add 200microl of alkaline lysis solution 2 , mix with inverting </span></li><li class="c4 c0"><span class="c3">Add 150 microl ice-cold alkaline lysis solution 3 mix with invertexing</span></li><li class="c4 c0"><span class="c3">Store tube on ice for 3-5 min</span></li><li class="c0 c4"><span class="c3">Centrifuge bacterial lysate ,5 min 13000rpm at 4deg in microfuge</span></li><li class="c4 c0"><span class="c3">Transfer supernatant to fresh tube</span></li><li class="c4 c0"><span class="c3">Add equal volume of phenol: chloroform</span></li><li class="c4 c0"><span class="c3">Mix the organic and aqueous phase by vortexing</span></li><li class="c4 c0"><span class="c3">Centrifuge at maximum speed 2 min at 4deg</span></li><li class="c4 c0"><span class="c3">Transfer the aqueous upper layer to fresh tube</span></li><li class="c4 c0"><span class="c3">Add 2 volume of ethanol</span></li><li class="c4 c0"><span class="c3">Mix by vortexing for 2 min</span></li><li class="c4 c0"><span class="c3">Centrifuge for 5min at 4deg</span></li><li class="c4 c0"><span class="c3">Remove supernatant by aspiration</span></li><li class="c4 c0"><span class="c3">Remove all liquid keeping inverted position</span></li><li class="c4 c0"><span class="c3">Add 1ml of ethanol to pellet</span></li><li class="c4 c0"><span class="c3">Centrifuge for 2min at 4deg</span></li><li class="c4 c0"><span class="c3">Remove all supernatant by aspiration</span></li><li class="c4 c0"><span class="c3">Remove ethanol beads and evaporate it at room temperature</span></li><li class="c4 c0"><span class="c3">Dissolve nucleic acid in 50microl of TE containing RNases A</span></li><li class="c4 c0"><span class="c3">Vortex the solution gently for 2 sec</span></li><li class="c4 c0"><span class="c3">Store DNA solution at -20deg</span></li></ol><p class="c0 c7 c22"><span></span></p><p class="c0 c22"><span>Result: no DNA pellet after precipitation by ethanol</span></p><p class="c0 c22"><span>Comment: Most probably step 12 is done in incorrect way</span></p><p class="c0 c7"><span></span></p><p class="c0"><span>Inoculation of a colony of BBa_K823016 and BBa_K1509003</span></p><p class="c0"><span>25/08/15</span></p><p class="c0"><span>Plasmid isolation of BBa_K1509003 using QIAGEN miniprep kit</span></p><p class="c0"><span>Protocol: QIAGEN Miniprep gravity column is used</span></p><p class="c0"><span>Result: No DNA after precipitation by isopropanol addition</span></p><p class="c0"><span>26/08/15</span></p><p class="c0"><span>Inoculation of BBa_K823016 and BBa_K1509003</span></p><p class="c0"><span>Plasmid isolation using QIAGEN Miniprep spin column</span></p><p class="c0"><span>Protocol: QIAGEN Miniprep spin column protocol is used</span></p><p class="c0"><span>Results:</span></p><a href="#" name="03314e51f11360a9d2dbab1bd0e6946dba809890"></a><a href="#" name="1"></a><table cellpadding="0" cellspacing="0" class="c2"><tbody><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">part</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">Concentration(ng/&micro;l)</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K823016</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">&nbsp;52.2</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K1509003</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">38.7</span></p></td></tr></tbody></table><p class="c0 c7"><span></span></p><p class="c0"><span>Inoculation of BBa_K823016 and BBa_K1509003 for alkaline lysis</span></p><p class="c0 c7"><span></span></p><p class="c0 c7"><span></span></p><p class="c0"><span>27/08/15</span></p><p class="c0"><span>Plasmid isolation using Alkaline lysis Miniprep in AB Lab</span></p><p class="c0"><span>Chemicals Required: Alkaline lysis buffer 1, 2, 3 buffer 2 freshly prepared, RNases</span></p><p class="c0"><span>Protocol: standard protocol is used from sambrook molecular cloning book as mentioned earlier </span></p><p class="c0"><span>Result </span></p><a href="#" name="2b7878fd2b0f2db50e7f91d80818952e3feebe23"></a><a href="#" name="2"></a><table cellpadding="0" cellspacing="0" class="c2"><tbody><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">part</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">Concentration(ng/&micro;l)</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K823016</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">2650.8</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K1509003</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">4080.3</span></p></td></tr></tbody></table><p class="c0 c7"><span></span></p><p class="c0"><span>This has high RNA contamination</span></p><p class="c0"><span>Gel Electrophoresis is done which shows high contamination of RNA</span></p><p class="c0 c7"><span></span></p><p class="c0"><span>2/09/15</span></p><p class="c0"><span>Inoculation of both BBa_K823016 and BBa_K1509003</span></p><p class="c0"><span>Plasmid isolation using QIAGEN Miniprep spin column</span></p><p class="c0"><span>Protocol: QIAGEN Miniprep spin column protocol is used</span></p><p class="c0"><span>Result:</span></p><p class="c0"><span>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; </span></p><a href="#" name="f34654009db90332628097eeff4e726342e11203"></a><a href="#" name="3"></a><table cellpadding="0" cellspacing="0" class="c2"><tbody><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">part</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">Concentration(ng/&micro;l)</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K823016</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">&nbsp;275.2</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K1509003</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">87.3</span></p></td></tr></tbody></table><p class="c0 c7"><span></span></p><p class="c0"><span>Gel Electrophoresis is done to check isolated plasmid</span></p><p class="c0"><span>8/09/15</span></p><p class="c0"><span>Inoculation of BBa_K1509003 in 5ml LB and 5&micro;l cam</span></p><p class="c0 c7"><span></span></p><p class="c0 c7"><span></span></p><p class="c0"><span>9/09/15</span></p><p class="c0"><span>Gel Electrophoresis (1%)</span></p><p class="c0"><span>Chemicals required: 0.5gm agarose, 50ml TAE, 5&micro;l EtBr</span></p><p class="c0"><span>Inoculation of both BBa_K823016 and BBa_K1509003 in 5ml LB each</span></p><p class="c0"><span>10/09/15</span></p><p class="c0"><span>Plasmid isolation using QIAGEN Miniprep spin column</span></p><p class="c0"><span>Protocol: QIAGEN Miniprep spin column protocol is used</span></p><p class="c0"><span>Result:</span></p><p class="c0"><span>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; </span></p><a href="#" name="aeeef95bed9e44d42ba9ef6b7bac92bdf75790b6"></a><a href="#" name="4"></a><table cellpadding="0" cellspacing="0" class="c2"><tbody><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">part</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">Concentration(ng/&micro;l)</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K823016</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">&nbsp;735</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K1509003</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">234</span></p></td></tr></tbody></table><p class="c0 c7"><span></span></p><p class="c0"><span>Single cut digestion</span></p><p class="c0"><span>BBa_K823016: 1&micro;l</span></p><p class="c0"><span>BBa_K1509003:3&micro;l</span></p><a href="#" name="7b1e6de2d1117931e0823fc80aba637223a503af"></a><a href="#" name="5"></a><table cellpadding="0" cellspacing="0" class="c18"><tbody><tr class="c6"><td class="c11" colspan="1" rowspan="1"><p class="c1 c0 c7"><span class="c5 c3"></span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K823016 (&micro;l)</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K1509003 (&micro;l)</span></p></td></tr><tr class="c6"><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">DNA</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">1</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">3</span></p></td></tr><tr class="c6"><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">ECoR1 buffer</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">1</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">1</span></p></td></tr><tr class="c6"><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">ECoR1 enzyme</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td></tr><tr class="c6"><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span>dH</span><span class="c13">2</span><span class="c5 c3">O</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">7.5</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">5.5</span></p></td></tr><tr class="c6"><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">Total </span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">10</span></p></td><td class="c11" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">10</span></p></td></tr></tbody></table><p class="c0 c7"><span></span></p><p class="c0"><span>Gel Electrophoresis: Fig1</span><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 276.00px; height: 244.00px;"><img alt="" src="images/image00.png" style="width: 276.00px; height: 244.00px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></span></p><p class="c0"><span>12/09/15</span></p><p class="c0"><span>&nbsp;Fig2</span></p><p class="c0"><span>Confirmation of plasmid isolated</span></p><p class="c0 c7"><span></span></p><p class="c0"><span>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; </span></p><p class="c0"><span>12/09/15</span></p><p class="c0"><span>Digestion for 3A assembly</span></p><p class="c0"><span>pSB1K3 which is a Kan backbone &nbsp;used for 3A assembly</span></p><p class="c0 c7"><span></span></p><p class="c0 c7"><span></span></p><a href="#" name="366d85563e792d67cfcbafbc92ff3b5233150491"></a><a href="#" name="6"></a><table cellpadding="0" cellspacing="0" class="c18"><tbody><tr class="c6"><td class="c17" colspan="1" rowspan="1"><p class="c1 c0 c7"><span class="c5 c3"></span></p></td><td class="c9" colspan="1" rowspan="1"><p class="c1 c0 c7"><span class="c5 c3"></span></p></td><td class="c20" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K823016 (&micro;l)</span></p></td><td class="c12" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K1509003 (&micro;l)</span></p></td></tr><tr class="c6"><td class="c17" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">1</span></p></td><td class="c9" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">NEB buffer 2</span></p></td><td class="c20" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">5</span></p></td><td class="c12" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">5</span></p></td></tr><tr class="c6"><td class="c17" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">2</span></p></td><td class="c9" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BSA</span></p></td><td class="c20" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td><td class="c12" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td></tr><tr class="c6"><td class="c17" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">3</span></p></td><td class="c9" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">EcoR1-HF</span></p></td><td class="c20" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td><td class="c12" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td></tr><tr class="c6"><td class="c17" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">4</span></p></td><td class="c9" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">Spel</span></p></td><td class="c20" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td><td class="c12" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">0.5</span></p></td></tr><tr class="c6"><td class="c17" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">5</span></p></td><td class="c9" colspan="1" rowspan="1"><p class="c1 c0"><span>dH</span><span class="c13">2</span><span class="c5 c3">0</span></p></td><td class="c20" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">18.5</span></p></td><td class="c12" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">18.5</span></p></td></tr><tr class="c6"><td class="c17" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">6</span></p></td><td class="c9" colspan="1" rowspan="1"><p class="c1 c0"><span class="c3 c5">Total</span></p></td><td class="c20" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">20</span></p></td><td class="c12" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">20</span></p></td></tr></tbody></table><p class="c0 c7"><span></span></p><p class="c0 c7"><span></span></p><p class="c0"><span>Protocol: standard iGEM 3A assembly protocol is used</span></p><p class="c0"><span>13/09/15</span></p><p class="c0"><span>Ligation using 3A assembly of biobricks</span></p><p class="c0"><span>Protocol: standard iGEM 3A assembly protocol is used</span></p><p class="c0"><span>15/09/15</span></p><p class="c0"><span>Transformation of ligated parts </span></p><p class="c0"><span>Protocol: IGEM transformation protocol is used</span></p><p class="c0"><span>Result: No colony is observed on both KAN media plates </span></p><p class="c0"><span>Comment: most probably failure of digestion</span></p><p class="c0"><span>16/09/15</span></p><p class="c0"><span>Inoculation of both BBa_K823016 and BBa_K1509003</span></p><p class="c0"><span>Plasmid isolation using QIAGEN Miniprep spin column</span></p><p class="c0"><span>Protocol: QIAGEN Miniprep spin column protocol is used</span></p><p class="c0"><span>Result:</span></p><p class="c0"><span>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; </span></p><a href="#" name="edd492d03c72482603f2fdab7a298129319088bf"></a><a href="#" name="7"></a><table cellpadding="0" cellspacing="0" class="c2"><tbody><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">part</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">Concentration(ng/&micro;l)</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K823016</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">&nbsp;102.3</span></p></td></tr><tr class="c6"><td class="c15" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">BBa_K1509003</span></p></td><td class="c8" colspan="1" rowspan="1"><p class="c1 c0"><span class="c5 c3">65.5</span></p></td></tr></tbody></table><p class="c0 c7"><span></span></p>
+</div>
+[<a href="top">top</a>]
 </body>
 </html>

Difference between revisions of "Team:IISER Pune/Notebook"

Latest revision as of 01:28, 19 September 2015

List of Note Books

Hijack Module (Fast robust oscillator) Notebook

Chromoproteins

Detection Enzyme Substrate Notebook