PCRs for fusions

Michał K.:

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

1-DNA ladder; 2-PCR product-translation fusion

Michał L., Ewa, Marcin:

1. Overnight inoculation and induction:

Top10 + 0,5 mM IPTG

pMPM-AID + Glc + 0,01% L-Ara

pTrc99a-AID + Glc + 0,5 mM IPTG