Team:Warsaw/Calendar-Main/3 October 2008

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Preparation of pSB2K3 + linker_alpha (BBa_K103009)

Piotr

  1. Transformation of TOP10 with ligation pSB2K3 + linker_alpha (BBa_K103009).
  2. Plating on LB with kanamycin.

Preparation of pSB2K3 + linker_omega (BBa_K103013)

Piotr

  1. Transformation of TOP10 with ligation pSB2K3 + linker_omega (BBa_K103013).
  2. Plating on LB with kanamycin.

Preparation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016)

Piotr

  1. Transformation of TOP10 with ligation pACYC177 + OmpA-linker-omega-linker (BBa_K103016).
  2. Plating on LB with kanamycin.

Preparation of BioBricks

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (pSB1A3+Omp_).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.
  3. Digest of pET15b+OmpA_omega with XbaI (Tango buffer). DNA ends blunting with Klenow fragment (3 hr).
  4. Gel electrophoresis and gel-out of proper band (pET15b+OmpA_omega without XbaI - 6500 bp).