Preparation of linker_alpha (BBa_K103009)

Michał K.

1. Colony PCR with LinLSXNE and AlphaPSpe primers on colonies from plates with transformations pSB1A3+ linker_alpha (BBa_K103009) (annealing temperature - 55°C,60 s of elongation step). No visible bands for proper product (Fig. 1).
2. Inoculation of some colonies which grown on plate - pSB2K3 + linker_alpha (BBa_K103009) to liquid LB + kanamycin.

Preparation of linker_omega (BBa_K103013)

Michał K.

Inoculation of some colonies which grown on plate - pSB2K3 + linker_omega (BBa_K103013) to liquid LB + kanamycin.

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

Michał K.

1. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations pACYC177 + OmpA-linker-omega-linker (BBa_K103016) (annealing temperature - 55°C,90 s of elongation step). No visible bands for proper product (Fig. 2).
2. Inoculation of some colonies which grown on plate transformation: pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to liquid LB + kanamycin.

Preparation of vector for pT7 constructs

Michał K.

Overnight selfligation of pET15b+OmpA_omega (with removed XbaI).

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Michał K.

Overnight ligation of isolated DNA fragments pSB2K3 (from 1 October) + OmpA_linker_omega_linker under Plac (BBa_K103018) (without EcoRI site).

Preparation of AID(BBa_K103001)

Michał K.

1. Digest of pSB1A3 plasmid with XbaI and PstI (Tango buffer). Dephosphorylation (CIAP) of plasmid.
2. Gel electrophoresis and gel-out of proper band - 2200 bp. Fig. 3.