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

Piotr, Weronika

1. Isolation of nazwa (Gfp genetrator), nazwa (RFP generator)
2. Digest of nazwa (Gfp genetrator), nazwa (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 with nazwa (Gfp genetrator), nazwa (RFP generator)
6. Chemotransformation of E.coli TOP10 with ligation products
7. Plating transformants on LB+Amp30+X-gal+IPTG

Clean-up of OmpA_alpha and OmpA_omega digest products.

Cloning PCL products on pKS vector

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.