Team:Paris Liliane Bettencourt/Notebook/2010/09/08/
From 2010.igem.org
Ligations (V + I)
- 1) pLux + LuxI
- 2) RBS/GFP/tt (E0240) + LuxI
- 3) Pms/mTet/LuxI + I13507/pLux/GFP
- 4) Ctrl pLux
- 5) Ctrl RBS/GFP/tt (E0240)
- 6) Ctrl Pms/mTet/LuxI
Transformation of 1-6 + positive control
Minipreps
- PmS-MTet-LuxI
- WTet
- MTet
- STet
- Ps
- pSB1C3 PmS-MTet-LuxI I
- pSB1C3 PsS-WTet-LuxI1 I
- PsS-WTet-LuxI2
- PsS-WTet-LuxI1
- pSB1C3 PsS-WTet-LuxI2 I
- pSB1C3 PmS-MTet-LuxI II
Restriction digest
With XbaI and PstI
- WTet
- MTet
- STet
With SpeI and PstI
- Ps
With EcoRI-HF and SpeI
- pSB1C3 PmS-MTet-LuxI I
- pSB1C3 PsS-WTet-LuxI1 I
- pSB1C3 PsS-WTet-LuxI2 I
- pSB1C3 PmS-MTet-LuxI II
- PmS-MTet-LuxI
Gel electrophoresis
- WTet
- MTet
- STet
- pSB1C3 PmS-MTet-LuxI I
- pSB1C3 PsS-WTet-LuxI1 I
- pSB1C3 PsS-WTet-LuxI2 I
- pSB1C3 PmS-MTet-LuxI II
- PmS-MTet-LuxI
Gel extraction
- WTet
- MTet
- STet
PCR purification
- Ps
Transformation
- PsS-WTet + pBad/Int
- PmS-MTet + pBad/Int
- positive control
Overnight cultures
- PmS-MTet-LuxI
- PsS-WTet-LuxI
- PmS Chr
- LuxI
- attP
- PsS-RFP
- PsW-RFP
- PsM-RFP
- PmS-RFP
- pBad/Int
From transformations of this morning :
- Final PmS-MTet x5
- LuxI/GFP x1
- pLux/LuxI x2 -> one colony didnt' grow