Preparation of linker_alpha (BBa_K103009)

Michał K., Piotr

1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009)).
2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
3. Inoculation of few more colonies which grown on plate with transformation: pSB2K3 + linker_alpha (BBa_K103009) to liquid LB + kanamycin.

Preparation of linker_omega (BBa_K103013)

Michał K., Piotr

1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_omega (BBa_K103013)).
2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/7_October_2008#fig1">Fig. 1.
3. Inoculation of few more colonies which grown on plates with transformation: pSB2K3 + linker_omega (BBa_K103013) to liquid LB + kanamycin.

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

Michał K., Piotr

1. Isolation of plasmid from culture inoculated on previous day (pACYC177 + OmpA-linker-omega-linker (BBa_K103016)).
2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/7_October_2008#fig2">Fig. 2.
3. Inoculation of few more colonies which grown on plate with transformation: pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to liquid LB + kanamycin.

Preparation of vector for pT7 constructs

Piotr

1. Transformation of TOP10 with selfligation of pET15b+OmpA_omega (with removed XbaI site).
2. Plating on LB with ampicillin.

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Piotr

1. Transformation of TOP10 with ligations pSB2K3 + BBa_K103018 (without internal EcoRI site).
2. Plating on LB with kanamycin.

Preparation of AID(BBa_K103001)

Michał K., Piotr

1. Ligation of DNA fragments: pSB1A3+ AID(BBa_K103001) (from 1 October).
2. Transformation of TOP10 with above ligation.
3. Plating on LB with ampicillin.

Preparation of AID under pBAD/araC (BBa_K103002)

Piotr

1. Digest of pMPMT5+AID with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment.
2. Overnight ligation of pMPMT5+AID to remove EcoRI site.