Preparation of 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 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 Fig. 1.

Preparation of 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 vector for pT7 constructs

Michał K.

Inoculation of colonies from plate with ligation of pET15b+OmpA_omega (with removed XbaI site) to liquid LB + ampicillin.

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Piotr

Inoculation of colonies from plate with ligation of pSB2K3 + BBa_K103018 (without internal EcoRI site) to liquid LB + kanamycin.

Preparation of AID(BBa_K103001)

Michał K.

1. Colony PCR with AIDlNrH and AIDpLinB Fig. 2 primers on colonies from plates with transformations pSB1A3+AID(BBa_K103001) (annealing temperature - 55°C, 60 s of elongation step).
2. Inoculation of confirmed colonies to liquid LB + ampicillin.

Preparation of AID under pBAD/araC (BBa_K103002)

Piotr

1. Transformation of TOP10 with ligation pMPMT5+AID (with removed EcoRI site).
2. Tranformants plating on LB with tetracycline.