Preparation of BioBricks

Michał K.

1. Isolation of plasmid from culture inoculated on previous day (RFP(?????)+ AID and pACYC177 + pET15b+OmpA_omega without XbaI).
2. Control digest of isolated RFP(?????)+ AID plasmids with EcoRI and PstI (Orange buffer) and pET15b+OmpA_omega without XbaI plasmids with XbaI and BamHI (Tango buffer). Gel electrophoresis - proper clones found for both ligations.
3. Digest of pET15b+OmpA_omega without XbaI plasmid with NdeI and SacI (BamHI buffer), dephosphorylation with CIAP
4. Gel electrophoresis and gel-out of proper band: ??????.

Piotr

1. Isolation of plasmid from culture inoculated on previous day (3kb??????) + pLac_OmpA_omega (without EcoRI site).
2. Control digest of isolated (3kb??????) + pLac_OmpA_omega with EcoRI and PstI (Orange buffer) proper clones found.
3. Inoculation of colonies from plate with ligation of pMPMT5+AID without EcoRI site to liquid LB + tetracycline.