Team:Paris/31 July 2009
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Revision as of 13:21, 10 August 2009 by Christophe.R (Talk | contribs)
NoteBook
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Lab work
DID dying to DH5a and MG4
We don't see florescence
->do it again but wash the cells before the DID
Miniprep
Miniprep (Pure Yield Tm Plasmid Miniprep System) for [S7] which contain g3p expression plasmid [Protocol]
Culture ON for glycerol stock
Biobrick resuspention
- 1/6I named 1I6 (BBa_R0040, TetR repressible promoter). Resistant to Ampicilin.
- 1/14N named 1N14 (BBa_10500, pBad/araC). Resistant to Kanamycin.
- 1/2J named 1J2 (BBa_J32015, PelB leader sequence). Resistant to Kanamycin.
- 1/18C named 1C18 (BBa_J61002, plasmid backbone). Resistant to Ampicilin.
- 1/7C named 1C7 (pSB2K3). Resistant to Kanamycin.
- 2/11P named 2P11 (BBa_I14032, pLac). Resistant to Kanamycin.
- 2/24C named 2C24 (BBa_B0014). Resistant to Kanamycin.
- 2/22P named 2p22 (BBa_K103006, OmpA linker). Resistant to Ampicilin.
- pFP141 (&DeltaN-Syn1A). Resistant to Kanamycin.
- pJM72 (His6-SNAP25). Resistant to Ampicilin.
- pTW2 (VAMP2-HIS6). Resistant to Ampicilin.
Transformation of precited Biobrick
- 250µl of DH5α competent bacteria (by RbCl)
- Add 2µl of DNA (Biobrick)
- Incubation during 20min on ice
- Heat choc 45second at 42°C
- Leave on ice 2min
- Add 1ml of LB preheated at 42°C
- Incubation 1 hour at 37°C under agitation
- Put 100µl of culture under plate
- Centrifuge 2min at 6rpm MAX for 2 min
- Put 100µl of concentrated culture
- Incubate overnight at 37°C
Transformation efficiency (cfu/µg)
- cfu/µg = nb colonies/µg DNA
- For RbCl, transformation efficiency = 1,2.107cfu/µg
To do list
Matricule | TODO |
Luc | SNARE Construction |
Romain | Design oligo Fec A |
Charlotte | Absent |
Stoff | Dev / Labs |
Chris | Global Design / Wiki / 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 |
Pierre | Modeling/SNARE |
Sylvain | Absent |
Guillaume | Labs / Protocol for g3p control / Thematique A / Sequencage OmpA-linker-G3P oligo |