Preparation of pMPMT5+T7 construct

Piotr

1. Isolation of hypothetical pMPMT5 with T7 RNA polymerase.
2. Control digest of pMPMT5 + T7 RNA-polymerase with EcoRI.
3. Gel electrophoresis.
4. Choice of proper clone and preparation for sequencing.
5. Design of primers for sequencing β-galactosidase from pZC320.
   Primers: pZCseqL and pZCseqP

Preparation of pMPMT5-AID+AIDT7 construct

Michał K.

Inoculation of 5 colonies of transformants (pMPMT5-AID with AID-T7 RNA-polymerase translation fusion) into liquid LB+tetracycline.

Blue/white and rifampicin test

Michał L., Ewa, Marcin:

1. Inoculation of of E. coli TOP10 with pZC320 into liquid LB broth (with 30 μg/ml of ampicillin).