Preparation of linker_alpha (BBa_K103009)

Michał K.

Overnight ligation of isolated DNA fragments: pSB2K3 + linker_alpha (BBa_K103009).

Preparation of linker_omega (BBa_K103013)

Michał K.

Overnight ligation of isolated DNA fragments: pSB2K3 + linker_omega (BBa_K103013).

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

Michał K.

Overnight ligation of DNA fragments isolated on 30 September : pACYC177 + OmpA-linker-omega-linker (BBa_K103016).

Preparation of OmpA-linker (BBa_K103006)

Michał K.

1. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations pSB1A3+OmpA-linker (BBa_K103006) (annealing temperature - 55°C,45 s of elongation step).
2. Gel electrophoresis( Fig. 1.Colony PCR with OmpaL_N and OmpaP_link.
   1. Marker
   4, 14, 19, 20, 24. Transformation confirmed.
   

   Preparation of Z(BBa_K103004)

   Michał K.

   1. Isolation of plasmids from culture inoculated on previous day (pSB1A3+Z(BBa_K103004)).
   2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found (
   3. Gel electrophoresis( Fig. 2.Control digest of pSB1A3+Z(BBa_K103004)
      1. Marker
      3. Clone carrying BBa_K103004

      Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

      Michał K.

      1. Digest of BBa_K103018 fragment with EcoRI and BcuI (BamHI buffer).
      2. Gel electrophoresis
      3. Gel electrophoresis (gel-out of proper band - 1200 bp.
      Fig. 3.Digest of OmpA_linker_omega_linker under Plac (BBa_K103018).
      1. Marker
      2. OmpA_linker_omega_linker under Plac (BBa_K103018)