Team:Warsaw/Calendar-Main/3 October 2008

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Preparation of linker_alpha (BBa_K103009)

Piotr

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

Preparation of linker_omega (BBa_K103013)

Piotr

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

Preparation of 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 OmpA-linker (BBa_K103006)

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (pSB1A3+OmpA-linker (BBa_K103006)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found (Fig. 1).
Fig. 1.Control digest of pSB1A3+OmpA-linker(BBa_K103006)
1. Marker
2-5. EcoRI/PstI control digest pSB1A3 carrying OmpA-linker (BBa_K103006)

Preparation of vector for pT7 constructs

Michał K.

  1. Digest of pET15b+OmpA_omega with XbaI (Tango buffer). DNA ends blunting with Klenow fragment (3 hr).
  2. Gel electrophoresis and gel-out of proper band (pET15b+OmpA_omega (with removed XbaI site) - 6500 bp). Fig. 2.
Fig. 2.XbaI digest of pET15b+OmpA_omega
1. Marker
2. XbaI digest pET15b+OmpA_omega


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