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

Piotr, Weronika

1. Isolation of pSB1A2 standard plasmids carrying parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator).
2. Digest of pSB1A2 standard plasmids carrying parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator) with NotI.
3. Digest of pZC320 with NotI and dephosphorylation with CIAP.
4. Gel electrophoresis and gel-out of proper bands.
5. Ligation of pZC320 with standard parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator).
6. Chemotransformation of E.coli TOP10 with ligation products.
7. Plating transformants on LB+Amp30+X-gal+IPTG.

Michał K.

Clean-up of OmpA_alpha and OmpA_omega digest products.

Cloning alpha-A and omega-A fusions on pKS

Michał L., Ewa, Marcin

1. Gel electophoresis of PCL products (Fig. 1).
2. Gel-out of proper bands (alpha-A: 1100 bp and omega-A: 750 bp).
3. Restriction digest of pKS vector and the PCL products with NotI and SacI (BamHI buffer) for 2 hours.
4. Ligation 1 hour.
5. Transformation of Top10 strain and screening on Amp₁₀₀ plates.