Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.

1. PCR - translational fusion: AID + T7 RNA-polymerase
   Primers:
   
   • AIDlNrH
   • T7pXbSal
   Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translational fusion
   Annealing temperature : 73°C
   Elongation time: 4 minutes
   20 cycles
   
2. Gel electrophoresis and isolation of proper band (3300 bp). Fig. 1.
3. Electrophoresis to estimate the concentration of isolated DNA.

Rifampicin test

Michał L., Ewa, Marcin

Overnight culture of the following strains:
Top10
Top10 + 0.5 mM IPTG
and Top10 carrying:
pMPM-T5-AID + Glc
pTrc99a-AID + Glc
pMPM-T5-AID + 0.01% L-Ara
pTrc99a-AID + 0.5 mM IPTG
Fig. 1. 1-DNA ladder; 2-PCR product-translational fusion