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

Michał K.

1. Isolation of plasmids from cultures inoculated on previous day pACYC177 + OmpA-linker-omega-linker (BBa_K103016).
2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.

Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K., Piotr

1. Gel electrophoresis of over night digest (pSB1A3 carrying alpha_linker under PT7 (BBa_K103019)) and gel-out of proper band - 800 bp.
2. Ligation of isolated DNA fragments: pSB2K3 (from 30 September) + alpha_linker under PT7 (BBa_K103019).
3. Transformation of TOP10 with above ligation.
4. Plating on LB with kanamycin.

Preparation of AraC+pBAD+AID

Piotr

1. Transformation of TOP10 with overnight ligation - pSB1A3 + AraC+pBAD+AID.
2. Planting on LB with ampicillin.

Piotr

1. Transformation of TOP10 with overnight ligation - RFP(?????) + AraC+pBAD+AID and 5kb(?????) + pT7_alpha.
2. Planting: RFP(?????) + AraC+pBAD+AID on LB with ampicillin and 5kb(?????) + alpha_linker under PT7 (BBa_K103019) on LB with kanamycin.