Change of the reporter from pZC320 with B-galactosidase to GFP or RFP

Piotr, Weronika

1. Isolation of plasmids from transformants.
2. Control digest of plasmids with NotI.
3. Gel electrophoresis of digested DNA - there where no proper plasmids.
4. Isolation of pZC320.
5. Isolation of pSB1A2 standard plasmids carrying parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator).
6. Overnight digest of pSB1A2 standard plasmids carrying parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator) with NotI.
7. Overnight digest of pZC320 with NotI and dephosphorylation with CIAP.

Cloning alpha-A and omega-A fusions on pKS

Michał L., Ewa, Marcin

1. Isolation of plasmids from transformants.
2. Control digest of plasmids with SacI+NotI (BamHI buffer).
3. Gel electrophoresis of digested DNA.

We have successfully cloned A-alpha and A-omega fusions on pKS vector. The DNA will be now sequenced to ensure that our constructs are correct.

Preparation of constructs on pET15b: OmpA_alpha and OmpA_omega

Paweł

1. Isolation of pET15b plasmid.
2. Overnight digest of pET15b plasmid with NdeI and BamHI (Tango 2x buffer), dephosphorylation with CIAP.