Team:Paris/5 August 2009

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Revision as of 19:44, 5 August 2009

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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 5th

Brain work

edit...

Lab work

FM4-64 dying (wash then stain)
n°1	n°2	n°3	n°4
500µl cells	-	-	-
centri 4000rpm 3min	-	-	-
resuspension 500µl MgSO4	-	-	-
1µl de colorant	1µl de colorant	2µl	2µl
incubation 30min 37°C	incubation 60min 37°C	incubation 30min 37°C	incubation 60min 37°C
depot sur lame	-	-	-
nothing	all cell fluo	so much dye	so much dye

FM4-64 dying (stain then wash)
n°5	n°6	n°9	n°10	n°13	n°14
500µl cells	-	-	-	500µl MgSO4	500µl Medium
1µl de colorant	1µl de colorant	1µl	1µl	1µl	1µl
incubation 30min 37°C	incubation 60min 37°C	incubation 30min 37°C	incubation 60min 37°C	incubation 60min 37°C	incubation 60min 37°C
centri 4000rpm 3min	-	-	-
resuspension 500µl MgSO4	resuspension 500µl MgSO4	+ filtrage	+ filtrage
depot sur lame	-	-	-	-	-
nothing	some cell fluo	noise	noise	noise	noise

Glycerol Stock

[S20] JW4251, [S21] JN0729, [S22] JN0728, [S23] JN0727, [S24] JN0940, [S24] JN0940, [S25] JW5100, [S25] JW5100, [S26] JN5181, [S27] FF64, [S28] NEC280.

Miniprep

Miniprep for P1/2/3/4/5/7/8
Same protocol as yesterday. Storage at -20°C.

PCR

Dilution oligo 1/10 (100µM to 10µM)
- 2µl Oligo
- 18µL H₂0 RNAse/DNAse free (Gibco)
Dilution DNA 1/10
- 2µl Genomic DNA of K12 MG1655 ΔrecA::Kan [F4]
- 18µL H₂0 RNAse/DNAse free (Gibco)
Mix x1 Vf=50µl in PCR tube (in ice) (Add with this order)
- 32,5µl H₂0 RNAse/DNAse free (Gibco)
- 10µl Buffer Polymerase fusion 5x
- 1µl dNTP
- 2,5µl Oligo F 10µM
- 2,5µl Oligo R 10µM
- 1µl Genomic DNA of K12 1/10
- 0,5µl Polymerase Fusion

Mix x6,5
- 211,25µl H₂0 RNAse/DNAse free (Gibco)
- 65µl Buffer Polymerase fusion 5x
- 6,5µl dNTP
- 6,5µl Genomic DNA of K12 1/10
- Put 44,5µl of Mix x6,5 in a PCR tube
- Add 2,5µl Oligo F 10µM
- Add 2,5µl Oligo R 10µM
- Add 0,5µl Polymerase Fusion

For A1/2/3/4/5/6, Program PHUSION:
- 1: 98°C, 1min
- 2: 98°C, 10sec
- 3: 65°C, 30sec
- 4: 72°C, 1min
- Goto 2 29x
- 72°C, 10min
- 4°C, -

Gel migration

Gel 1%: 5µl of [A1|A2|100bp|A3|A4|1kb|A5|A6]
- A1 = Amplification of N-term domain of M13's g3p = 274bp (Useless because we don't use g3p plasmide as matrice => Ctrl-)
- A2 = Amplification of N-term domain of colicin E3 = 1036bp (Useless because we don't use the right matrice => Ctrl-)
- A3 = Amplification of clyA with RBS in the primer = 987bp
- A4 = Amplification of OmpA signal for export to periplasm = 135bp
- A5 = Amplification of tolR = 279bp (Try again because of the second band)
- A6 = Amplification of tatA with RBS = 1634bp (Try again because of the second band)

ON culture

[S8] DH5α(pSB2K3): LB Kan
[29] DH5α(pSB1A3): LB Amp
[30] S113 iGEM08 = DH5α(BBa_K136050): LB Amp

To do list

Matricule	TODO
Luc	bacteria coloration microscope protocol
Romain	Look for his feet
Charlotte	bibliography on SNAREs/Autotransporters/Jun and Fos complex
Stoff	merge algo protocol/ Wiki
Chris	modeling: TeTR
Lisa	WTF !!!
Caroline	bacteria coloration microscope protocol
Souf	if(!oligo){return Wiki;} return PCR;
Vicard	Lab : Gel / purification / insers
Pierre	Alice / Tol/Pal modelisation / geophysics
Sylvain	if(!oligo){return Maltose;} return PCR;
Guillaume	Miniprep for P(1/2/3/4/5/7/8) / New P7 digestion? / Glycerol Stock (DH5α/Kaio strains)

@@ Line 157: / Line 157: @@
 PCR
 </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px">
-*Dilution oligo 1/10
+*Dilution oligo 1/10 (100µM to 10µM)
 **2µl Oligo
 **18µL H<sub>2</sub>0 RNAse/DNAse free (Gibco)
 *Dilution DNA 1/10
-**2µl Genomic DNA of K12 MG1655 &Delta;recA::Kan [[https://2009.igem.org/Team:Paris/Fridge | F4]]
+**2µl Genomic DNA of K12 MG1655 &Delta;recA::Kan [[https://2009.igem.org/Team:Paris/Fridge F4]]
 **18µL H<sub>2</sub>0 RNAse/DNAse free (Gibco)
+*Mix x1 Vf=50µl in PCR tube (in ice) (Add with this order)
+**32,5µl H<sub>2</sub>0 RNAse/DNAse free (Gibco)
+**10µl Buffer Polymerase fusion 5x
+**1µl dNTP
+**2,5µl Oligo F 10µM
+**2,5µl Oligo R 10µM
+**1µl Genomic DNA of K12 1/10
+**0,5µl Polymerase Fusion
+<br>
+*Mix x6,5
+**211,25µl H<sub>2</sub>0 RNAse/DNAse free (Gibco)
+**65µl Buffer Polymerase fusion 5x
+**6,5µl dNTP
+**6,5µl Genomic DNA of K12 1/10
+**Put 44,5µl of Mix x6,5 in a PCR tube
+**Add 2,5µl Oligo F 10µM
+**Add 2,5µl Oligo R 10µM
+**Add 0,5µl Polymerase Fusion
+<br>
+*For A1/2/3/4/5/6, Program PHUSION:
+**1: 98°C, 1min
+**2: 98°C, 10sec
+**3: 65°C, 30sec
+**4: 72°C, 1min
+**Goto 2 29x
+**72°C, 10min
+**4°C, -
 </div>
 <div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px">

Team:Paris/5 August 2009

From 2009.igem.org

Revision as of 19:44, 5 August 2009

Contents

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Brain work

Lab work

To do list