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Team:Paris/5 August 2009
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PCR | PCR | ||
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| + | *Dilution oligo 1/10 | ||
| + | **2µl Oligo | ||
| + | **18µL H<sub>2</sub>0 RNAse/DNAse free (Gibco) | ||
| + | *Dilution DNA 1/10 | ||
| + | **2µl Genomic DNA of K12 MG1655 ΔrecA::Kan [[https://2009.igem.org/Team:Paris/Fridge | F4]] | ||
| + | **18µL H<sub>2</sub>0 RNAse/DNAse free (Gibco) | ||
| + | |||
| + | |||
</div> | </div> | ||
Revision as of 19:34, 5 August 2009
Contents |
NoteBook
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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
- 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)
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) |
