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.
3. Confirmed transformant colonies inoculated to liquid LB with ampicillin.

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.

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Michał K.

1. Digest of pLac_OmpA_omega fragment with EcoRI and BcuI (BamHI buffer).
2. Gel elctrophoresis and gel-out of proper band - 1200 bp.