Team:Paris/5 August 2009
From 2009.igem.org
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- | + | Glycerol Stock | |
</div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
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- | + | Miniprep | |
</div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
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- | + | PCR | |
</div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
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- | + | Gel migration | |
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- | + | *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) | ||
+ | <center>[[Image:100bp.png]][[Image:1kb.gif]][[Image:PCR_050809.png]]</center> | ||
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- | + | ON culture | |
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- | + | *[S8] DH5α(pSB2K3): LB Kan | |
+ | *[29] DH5α(pSB1A3): LB Amp | ||
+ | *[30] S113 iGEM08 = DH5α(BBa_K136050): LB Amp | ||
</div> | </div> | ||
<html> | <html> |
Revision as of 19:12, 5 August 2009
Contents |
NoteBook
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Brain work
edit...
Lab work
FM4-64 dying (wash then stain) | ||||||||||
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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
Miniprep
PCR
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) |