Team:Warsaw/Calendar-Main/16 October 2008

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Preparation of pSB2K3 + _alpha

Michał K.

  • Isolation of plasmids from culture inoculated on previous day.
  • 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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