INSA-Lyon/12 October 2010
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October 12th
Liquid cultures of 3 clones from the ligation 18A/OmpR234 of the 10/10
Digestion :
- 12/13 : with E+X
- pha C with E+S
- phasin-intein, 1N : with S+P
- 2P, L2/1N, phaC : with X+P
- L2/1N, curli : with E+P
- phaA/phaB : with E
Agar gel electrophoresis for verification and purification :
- 1N, L2/1N(E+P), curli and phaA/phaB : No DNA appear
- 12/13, pha C(X+P and E+S), phasin-intein, 2P, L2/1N (X+P) : OK purification
Ligation overnight at 15°C , two series one with purified DNA and the other with DNA without purification:
- 18A + L2/1N in Amp backbone
- Curli + phaC in Amp backbone
- phasin-intein/2P : in kan
- phaC + 12/13
- L2/1N + phaC
- L2/1N + phasin-intein
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