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).

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.