Team:Paris/30 July 2009
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- | == | + | ===Lab work=== |
+ | <div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px"> | ||
+ | Reception of strains | ||
+ | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
+ | *New DH5α | ||
+ | *DH5α with g3p in plasmid | ||
+ | </div> | ||
- | + | <div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px"> | |
+ | Glycerol stock of new DH5α [S6] | ||
+ | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
+ | *Stock in 20% glycerol at 80°C | ||
+ | </div> | ||
+ | <div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px"> | ||
+ | New competent DH5α by RbCl | ||
+ | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
+ | *Competent bacteria | ||
+ | **1/50 ON culture of New DH5α | ||
+ | **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) | ||
+ | </div> | ||
- | < | + | <div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px"> |
+ | Overnight culture for [S3] Δnlp1, [S4] TB28, [S5] MG4, [S6] DH5α, [S7] DH5α g3p | ||
+ | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
+ | *[https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S3 S3] Δnlp1: 5ml LB + 5µl Kan 1000x | ||
+ | *[https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S4 S4] TB28 : 5ml LB | ||
+ | *[https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S5 S5] MG4: 5ml LB + 5µl Kan 1000x + 250µl NaCl 20% | ||
+ | *[https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S6 S6] DH5α: 5ml LB | ||
+ | *[https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S7 S7] DH5α g3p: 6ml LB + 6µl Amp 1000x | ||
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- | <div | + | |
+ | <div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px"> | ||
+ | Stock of NaCl 20% | ||
+ | </div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px"> | ||
+ | *100ml of H<sub>2</sub>O | ||
+ | *20g of NaCl | ||
+ | *Filtration | ||
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- | + | ===To do list=== | |
- | ==To do list== | + | |
- | + | ||
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{| | {| | ||
|- style="background: #ccccff; text-align: center;" | |- style="background: #ccccff; text-align: center;" | ||
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|Thematique A / Bacterial display / transfo ? / sequencage OmpA-linker-G3P oligo | |Thematique A / Bacterial display / transfo ? / sequencage OmpA-linker-G3P oligo | ||
|} | |} | ||
+ | |||
+ | |||
+ | {{Paris2009_Calendar_Link|29_July_2009|31_July_2009}} |
Latest revision as of 13:20, 10 August 2009
NoteBook
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Lab work
Reception of strains
- New DH5α
- DH5α with g3p in plasmid
Glycerol stock of new DH5α [S6]
- Stock in 20% glycerol at 80°C
New competent DH5α by RbCl
- Competent bacteria
- 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)
Overnight culture for [S3] Δnlp1, [S4] TB28, [S5] MG4, [S6] DH5α, [S7] DH5α g3p
Stock of NaCl 20%
- 100ml of H2O
- 20g of NaCl
- Filtration
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 |