Team:Paris/18 August 2009

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NoteBook

June
M	T	W	T	F	S	S
[http://2009.igem.org/Team:Paris/1_June_2009 1]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/2_June_2009&action=edit 2]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/3_June_2009&action=edit 3]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/4_June_2009&action=edit 4]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/5_June_2009&action=edit 5]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/6_June_2009&action=edit 6]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/7_June_2009&action=edit 7]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/8_June_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/9_June_2009&action=edit 9]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/10_June_2009&action=edit 10]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/11_June_2009&action=edit 11]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/12_June_2009&action=edit 12]	[http://2009.igem.org/Team:Paris/13_June_2009 13]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/14_June_2009&action=edit 14]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/15_June_2009&action=edit 15]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/16_June_2009&action=edit 16]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/17_June_2009&action=edit 17]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/18_June_2009&action=edit 18]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/19_June_2009&action=edit 19]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/20_June_2009&action=edit 20]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/21_June_2009&action=edit 21]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/22_June_2009&action=edit 22]	[http://2009.igem.org/Team:Paris/23_June_2009 23]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/24_June_2009&action=edit 24]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/25_June_2009&action=edit 25]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/26_June_2009&action=edit 26]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/27_June_2009&action=edit 27]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/28_June_2009&action=edit 28]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/29_June_2009&action=edit 29]	[http://2009.igem.org/Team:Paris/30_June_2009 30]

July
M	T	W	T	F	S	S
		[http://2009.igem.org/wiki/index.php?title=Team:Paris/1_July_2009&action=edit 1]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/2_July_2009&action=edit 2]	[http://2009.igem.org/Team:Paris/3_July_2009 3]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/4_July_2009&action=edit 4]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/5_July_2009&action=edit 5]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/6_July_2009&action=edit 6]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/7_July_2009&action=edit 7]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/8_July_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/9_July_2009&action=edit 9]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/10_July_2009&action=edit 10]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/11_July_2009&action=edit 11]	[http://2009.igem.org/wiki/index.php?title=Team:Paris/12_July_2009&action=edit 12]
[http://2009.igem.org/Team:Paris/13_July_2009 13]	[http://2009.igem.org/Team:Paris/14_July_2009 14]	[http://2009.igem.org/Team:Paris/15_July_2009 15]	[http://2009.igem.org/Team:Paris/16_July_2009 16]	[http://2009.igem.org/Team:Paris/17_July_2009 17]	[http://2009.igem.org/Team:Paris/18_July_2009 18]	[http://2009.igem.org/Team:Paris/19_July_2009 19]
[http://2009.igem.org/Team:Paris/20_July_2009 20]	[http://2009.igem.org/Team:Paris/21_July_2009 21]	[http://2009.igem.org/Team:Paris/22_July_2009 22]	[http://2009.igem.org/Team:Paris/23_July_2009 23]	[http://2009.igem.org/Team:Paris/24_July_2009 24]	[http://2009.igem.org/Team:Paris/25_July_2009 25]	[http://2009.igem.org/Team:Paris/26_July_2009 26]
[http://2009.igem.org/Team:Paris/27_July_2009 27]	[http://2009.igem.org/Team:Paris/28_July_2009 28]	[http://2009.igem.org/Team:Paris/29_July_2009 29]	[http://2009.igem.org/Team:Paris/30_July_2009 30]	[http://2009.igem.org/Team:Paris/31_July_2009 31]

August
M	T	W	T	F	S	S
					[http://2009.igem.org/Team:Paris/1_August_2009 1]	[http://2009.igem.org/Team:Paris/2_August_2009 2]
[http://2009.igem.org/Team:Paris/3_August_2009 3]	[http://2009.igem.org/Team:Paris/4_August_2009 4]	[http://2009.igem.org/Team:Paris/5_August_2009 5]	[http://2009.igem.org/Team:Paris/6_August_2009 6]	[http://2009.igem.org/Team:Paris/7_August_2009 7]	[http://2009.igem.org/Team:Paris/8_August_2009 8]	[http://2009.igem.org/Team:Paris/9_August_2009 9]
[http://2009.igem.org/Team:Paris/10_August_2009 10]	[http://2009.igem.org/Team:Paris/11_August_2009 11]	[http://2009.igem.org/Team:Paris/12_August_2009 12]	[http://2009.igem.org/Team:Paris/13_August_2009 13]	[http://2009.igem.org/Team:Paris/14_August_2009 14]	[http://2009.igem.org/Team:Paris/15_August_2009 15]	[http://2009.igem.org/Team:Paris/16_August_2009 16]
[http://2009.igem.org/Team:Paris/17_August_2009 17]	[http://2009.igem.org/Team:Paris/18_August_2009 18]	[http://2009.igem.org/Team:Paris/19_August_2009 19]	[http://2009.igem.org/Team:Paris/20_August_2009 20]	[http://2009.igem.org/Team:Paris/21_August_2009 21]	[http://2009.igem.org/Team:Paris/22_August_2009 22]	[http://2009.igem.org/Team:Paris/23_August_2009 23]
[http://2009.igem.org/Team:Paris/24_August_2009 24]	[http://2009.igem.org/Team:Paris/25_August_2009 25]	[http://2009.igem.org/Team:Paris/26_August_2009 26]	[http://2009.igem.org/Team:Paris/27_August_2009 27]	[http://2009.igem.org/Team:Paris/28_August_2009 28]	[http://2009.igem.org/Team:Paris/29_August_2009 29]	[http://2009.igem.org/Team:Paris/30_August_2009 30]
[http://2009.igem.org/Team:Paris/31_August_2009 31]

September
M	T	W	T	F	S	S
	[http://2009.igem.org/Team:Paris/1_September_2009 1]	[http://2009.igem.org/Team:Paris/2_September_2009 2]	[http://2009.igem.org/Team:Paris/3_September_2009 3]	[http://2009.igem.org/Team:Paris/4_September_2009 4]	[http://2009.igem.org/Team:Paris/5_September_2009 5]	[http://2009.igem.org/Team:Paris/6_September_2009 6]
[http://2009.igem.org/Team:Paris/7_September_2009 7]	[http://2009.igem.org/Team:Paris/8_September_2009 8]	[http://2009.igem.org/Team:Paris/9_September_2009 9]	[http://2009.igem.org/Team:Paris/10_September_2009 10]	[http://2009.igem.org/Team:Paris/11_September_2009 11]	[http://2009.igem.org/Team:Paris/12_September_2009 12]	[http://2009.igem.org/Team:Paris/13_September_2009 13]
[http://2009.igem.org/Team:Paris/14_September_2009 14]	[http://2009.igem.org/Team:Paris/15_September_2009 15]	[http://2009.igem.org/Team:Paris/16_September_2009 16]	[http://2009.igem.org/Team:Paris/17_September_2009 17]	[http://2009.igem.org/Team:Paris/18_September_2009 18]	[http://2009.igem.org/Team:Paris/19_September_2009 19]	[http://2009.igem.org/Team:Paris/20_September_2009 20]
[http://2009.igem.org/Team:Paris/21_September_2009 21]	[http://2009.igem.org/Team:Paris/22_September_2009 22]	[http://2009.igem.org/Team:Paris/23_September_2009 23]	[http://2009.igem.org/Team:Paris/24_September_2009 24]	[http://2009.igem.org/Team:Paris/25_September_2009 25]	[http://2009.igem.org/Team:Paris/26_September_2009 26]	[http://2009.igem.org/Team:Paris/27_September_2009 27]
[http://2009.igem.org/Team:Paris/28_September_2009 28]	[http://2009.igem.org/Team:Paris/29_September_2009 29]	[http://2009.igem.org/Team:Paris/30_September_2009 30]

October
M	T	W	T	F	S	S
			[http://2009.igem.org/Team:Paris/1_October_2009 1]	[http://2009.igem.org/Team:Paris/2_October_2009 2]	[http://2009.igem.org/Team:Paris/3_October_2009 3]	[http://2009.igem.org/Team:Paris/4_October_2009 4]
[http://2009.igem.org/Team:Paris/5_October_2009 5]	[http://2009.igem.org/Team:Paris/6_October_2009 6]	[http://2009.igem.org/Team:Paris/7_October_2009 7]	[http://2009.igem.org/Team:Paris/8_October_2009 8]	[http://2009.igem.org/Team:Paris/9_October_2009 9]	[http://2009.igem.org/Team:Paris/10_October_2009 10]	[http://2009.igem.org/Team:Paris/11_October_2009 11]
[http://2009.igem.org/Team:Paris/12_October_2009 12]	[http://2009.igem.org/Team:Paris/13_October_2009 13]	[http://2009.igem.org/Team:Paris/14_October_2009 14]	[http://2009.igem.org/Team:Paris/15_October_2009 15]	[http://2009.igem.org/Team:Paris/16_October_2009 16]	[http://2009.igem.org/Team:Paris/17_October_2009 17]	[http://2009.igem.org/Team:Paris/18_October_2009 18]
[http://2009.igem.org/Team:Paris/19_October_2009 19]	[http://2009.igem.org/Team:Paris/20_October_2009 20]	[http://2009.igem.org/Team:Paris/21_October_2009 21]	[http://2009.igem.org/Team:Paris/22_October_2009 22]	[http://2009.igem.org/Team:Paris/23_October_2009 23]	[http://2009.igem.org/Team:Paris/24_October_2009 24]	[http://2009.igem.org/Team:Paris/25_October_2009 25]
[http://2009.igem.org/Team:Paris/26_October_2009 26]	[http://2009.igem.org/Team:Paris/27_October_2009 27]	[http://2009.igem.org/Team:Paris/28_October_2009 28]	[http://2009.igem.org/Team:Paris/29_October_2009 29]	[http://2009.igem.org/Team:Paris/30_October_2009 30]	[http://2009.igem.org/Team:Paris/31_October_2009 31]

← Yesterday - Tomorrow →

August 18th

Lab work

PCR on colony w/ Soufifi

The ligation pLAC/RBS-TetR worked --> no clone on the negatif control and some clone on the "good" plates. PCR on colony is required to check the validity of our clones.

Mix : total volume = 25uL

Super Mix 2X: 12,5 uL

VF2 (10uM): 0,5 uL

VR (10 uM): 0,5 uL

H20 : 6,5 uL

diluted bacteria : 5 uL

Tm = 55°C

Elongation time = 1 min

size expected :

- pLac/RBS-TetR : +/- 700 bp

- VF2 and VR plasmid amplification : +/- 150 bp

- final size : +/- 850 bp

Digestion

The ligation that I have set up yesterday (pSB1A3 w/ D13 or D14 ) didn't work so I decided to redigest PCR products (A11 (RBS-ompA signal = D13) and A13 (TolRII = D14)).

Mix : total volume = 30 uL

For A11 (RBS-OmpA signal):

DNA = 15 uL

10X buffer: 3 uL

SpeI: 1 uL

PstI: 1 uL

BSA: 0,5 uL

H2O: 9,5 uL

For A13:

DNA = 10 uL

10X buffer: 3 uL

XbaI: 1 uL

PstI: 1 uL

BSA: 0,5 uL

H2O: 14,5 uL

Other digestions were done :

- The double terminator (BBa_0034) by EcoRI/XbaI and SpeI/PstI (same mix as the one for A13)

- The pTet (BBa_....) by SpeI/PstI and EcoRI/PstI

Gel purification w/ Bobode

Promega kit, 3 uL loadded

Gel purification

PCR on colonies:

using for 1 tube:

Master Mix 2X : 12,5µL

Oligo VF2 (10µM): 0,5µL

Oligo VR (10µM): 0,5µL

bacteria in H20 (see PCR on colonies protocol): 5µL

H20 : 6,5µL

PCR on colonies Check OK for colonie 7, 8, 9 and 10

Control+ (with PSB1A3)

Colonie 7, 8 and 10 OK.

Stock Glycerol

stock glycerol for colonie 3 ClyA in PSB1A3 using: -1/3 glycerol (333µL) + 2/3 bacteria (667µL)

Miniprep for colonie 3, 7, 8 and 10

← Yesterday - Tomorrow →