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

Piotr, Weronika

1. Inoculation of 10ml LB with E.coli TOP10 carrying pZC
2. Isolation and chemotransformation with pSB1A2 standard plasmids carrying parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator)
3. Plating of chemotransformants on LB+ampicillin

Preparation of constructs: OmpA_alpha and OmpA_omega

Michał K.

Repetition of PCRs and gel-out purifications from 16 June.

Preparation of alpha-A and omega-A fusions

Michał L. Marcin and Ewa

1. We've run gradient PCR to obtain Alpha-link and link-A.
   • DNA template: pDRIVE-TAPTAG
   • Taq polymerase
   • buffer suplemented with (NH₄)₂SO₄
   • MgCl₂ concentration gradient (from 1 to 4 mM)
   • annealing temp 55°C
   • elongation time 1 - 2 minutes
2. We have successfully amplified Alpha-link and link-A (Fig. 1). Now it's time for PCL (Polymerase Chain Ligation) to create fusions Alpha-A and Omega-A.