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.
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.
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 without XbaI.
2. Plating on LB with ampicillin.

Preparation of BioBricks

Michał K.

1. Overnight ligation of DNA fragments: pSB1A3+ AID.

Piotr

1. Transformation of TOP10 with ligations pSB2K3 + pLac_OmpA_omega (without EcoRI site) and pET15b+OmpA_omega without XbaI.
2. Plating: pSB2K3 + pLac_OmpA_omega (without EcoRI site) on LB with kanamycin and pET15b+OmpA_omega without XbaI on LB with ampicillin.
3. Digest of pMPMT5+AID with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment.
4. Overnight ligation of pMPMT5+AID without EcoRI site.