Team:Warsaw/Calendar-Main/16 October 2008

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Preparation of pSB2K3 + pSB2K3 + linker_alpha (BBa_K103009)

Michał K.

  1. Isolation of plasmids from culture inoculated on previous day.
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.

Preparation of BioBricks

Michał K.

  1. Isolation of plasmids from culture inoculated on previous day 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2 and RFP(?????) + AraC+pBAD+AID.
  2. Control digest of isolated plasmids: 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2, RFP(?????) + AraC+pBAD+AID and RFP(?????) + pT7_alpha (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found for RFP(?????) + AraC+pBAD+AID ligation but we found good clones for other ligations.
  3. Inoculation of colonies from plate with ligation of pACYC177 + OmpA_omega_ to LB with kanamycin.
  4. Repetition of overnight ligation of isolated DNA fragments: RFP(?????) + AraC+pBAD+AID.
  5. Overnight digest of RFP(?????) + pT7_alpha EcoRI and BcuI (BamHI buffer).

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