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 :

  1. 12/13 : with E+X
  2. pha C with E+S
  3. phasin-intein, 1N : with S+P
  4. 2P, L2/1N, phaC : with X+P
  5. L2/1N, curli : with E+P
  6. phaA/phaB : with E

Agar gel electrophoresis for verification and purification :

  1. 1N, L2/1N(E+P), curli and phaA/phaB : No DNA appear
  2. 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:

  1. 18A + L2/1N in Amp backbone
  2. Curli + phaC in Amp backbone
  3. phasin-intein/2P : in kan
  4. phaC + 12/13
  5. L2/1N + phaC
  6. L2/1N + phasin-intein








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