Team:Paris/30 July 2009
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==Lab work== | ==Lab work== | ||
- | '''Reception of strains:''' | + | *'''Reception of strains:''' |
- | *New DH5α | + | **New DH5α |
- | *DH5α with g3p in plasmid | + | **DH5α with g3p in plasmid |
+ | *'''Glycerol stock of new DH5α [S6]''' | ||
- | '''New DH5α | + | *'''New competent DH5α by RbCl''' |
- | *1/50 ON culture of New DH5α | + | **1/50 ON culture of New DH5α |
- | *OD<sub>600</sub>=0,6 | + | **OD<sub>600</sub>=0,6 |
+ | *'''Transformation with puc19 (10pg/µl)''' | ||
+ | **250µl of RbCl competent DH5α | ||
+ | **12,5µl puc19 | ||
+ | **Leave 20 min in ice | ||
+ | **45s at 42°C | ||
+ | **Leave 2 min in ice | ||
+ | **Add 1ml of hot LB (42°C) | ||
+ | **1h at 37 under agitation | ||
+ | **Put on plate (LB agar + Amp) | ||
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**[S3] Δnlp1: 5ml LB + 5µl Kan 1000x | **[S3] Δnlp1: 5ml LB + 5µl Kan 1000x | ||
**[S4] TB28 : 5ml LB | **[S4] TB28 : 5ml LB | ||
- | + | **[S5] MG4: 5ml LB + 5µl Kan 1000x + 250µl NaCl 20% | |
- | *[S6] DH5α: 5ml LB | + | **[S6] DH5α: 5ml LB |
**[S7] DH5α g3p: 6ml LB + 6µl Amp 1000x | **[S7] DH5α g3p: 6ml LB + 6µl Amp 1000x | ||
- | '''Stock of NaCL 10%''' [F11] | + | *'''Stock of NaCL 10%''' [F11] |
<html> | <html> |
Revision as of 01:17, 31 July 2009
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Brain work
edit please ^^
Lab work
- Reception of strains:
- New DH5α
- DH5α with g3p in plasmid
- Glycerol stock of new DH5α [S6]
- New competent DH5α by RbCl
- 1/50 ON culture of New DH5α
- OD600=0,6
- Transformation with puc19 (10pg/µl)
- 250µl of RbCl competent DH5α
- 12,5µl puc19
- Leave 20 min in ice
- 45s at 42°C
- Leave 2 min in ice
- Add 1ml of hot LB (42°C)
- 1h at 37 under agitation
- Put on plate (LB agar + Amp)
- ON culture for [S3] Δnlp1, [S4] TB28, [S5] MG4, [S6] DH5α, [S7] DH5α g3p
- [S3] Δnlp1: 5ml LB + 5µl Kan 1000x
- [S4] TB28 : 5ml LB
- [S5] MG4: 5ml LB + 5µl Kan 1000x + 250µl NaCl 20%
- [S6] DH5α: 5ml LB
- [S7] DH5α g3p: 6ml LB + 6µl Amp 1000x
- Stock of NaCL 10% [F11]
To do list
Matricule | TODO |
Luc | Sequence SNARE / Labs / Construction |
Romain | Design oligo Fec A |
Charlotte | Oligo Fec A /Global design / Meeting Sara |
Stoff | Dev / Labs |
Chris | Global Design / Meeting Sara / Modeling |
Lisa | Oligo Tol/Pal / Oligo Sequencing / Tol/Pal Protocol verification / Keio / Theme A writting |
Caroline | Oligo Tol/Pal / Oligo Sequencing / Tol/Pal Protocol verification / Keio / Theme A writting |
Souf | Tat/Sec system / Design signal peptid |
Vicard | ClyA protocol / using TAT ? |
Pierre | Modeling/SNARE |
Sylvain | Labs / Maltose (thomas) |
Guillaume | Thematique A / Bacterial display / transfo ? / sequencage OmpA-linker-G3P oligo |