Preparation of pSB2K3 + linker_alpha (BBa_K103009)

Piotr

1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009)).
2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.

Preparation of pSB2K3 + linker_omega (BBa_K103013)

Piotr

1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_omega (BBa_K103013)).
2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.

Preparation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016)

Piotr

1. Isolation of plasmid from culture 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 BioBricks

Michał K.

1. Colony PCR with AIDlNrH and AIDpLinB primers on colonies from plates with transformations pSB1A3+ AID (annealing temperature - 55°C, 60 s of elongation step).
2. Inoculation of confirmed colonies to liquid LB + ampicillin.
3. Inoculation of colonies from plate with ligation of pET15b+OmpA_omega without XbaI to liquid LB + ampicillin.

Piotr

1. Inoculation of colonies from plate with ligation of pSB2K3 + pLac_OmpA_omega (without EcoRI site) to liquid LB + kanamycin.
2. Transformation of TOP10 with ligation pMPMT5+AID without EcoRI site.
3. Tranformants planting on LB with tetracycline.