Team:Paris Liliane Bettencourt/Notebook/2010/09/09/
From 2010.igem.org
Minipreps
- PmS-MTet-LuxI
- LuxI
- attP
- PsS-RFP
- PsW-RFP
- PsM-RFP
- PmS-RFP
- pBad/Int
- Final PmS-MTet x5
- LuxI/GFP x1
- pLux/LuxI x1
Restriction digest
With XbaI and PstI
- LuxI/GFP
- I13507/pLux/GFP
- RBS-GFP-tt (E0240)
With SpeI and PstI
- PmS-MTet-LuxI
- LuxI
- pLux/LuxI
- pLux
With EcoRI-HF and PstI
- PmS-MTet-LuxI
- PsS-RFP
- PsW-RFP
- PsM-RFP
- PmS-RFP
- attP
- Final PmS-MTet x5
Gel electrophoresis
- PmS-MTet-LuxI (E+P)
- PsS-RFP
- PsW-RFP
- PsM-RFP
- PmS-RFP
- attP
- I13507/pLux/GFP
- Final PmS-MTet 1
- Final PmS-MTet 2
- Final PmS-MTet 3
- Final PmS-MTet 4
- Final PmS-MTet 5
- LuxI/GFP
- RBS-GFP-tt (E0240)
Ligations (V + I)
- 1) Ps + WTet
- 2) Ps + MTet
- 3) Ps + STet
- 4) Ctrl Ps
Make competent cells
- Turbocells (100mL of culture)
- PmS Chr (to transform with pBad/Int)
Transformation of 1-4 + positive control (pUC19) for new Turbocells
Overnight cultures
- Plambda Int
From transformations of yesterday :
- PsS-WTet-LuxI + pBad/Int x2
- PmS-MTet-LuxI + pBad/Int x 2