Preparation of pMPMT5-AID+AIDT7 construct

1. Digest of isolated PCR product (from previous day) and pMPMT5-AID with HindIII and XbaI, dephosphorylation of pMPMT5-AID with CIAP
2. Clean-up of digested PCR product.
3. Gel electrophoresis of digested pMPMT5-AID and gel-out of proper band (4800 bp)
4. Overnight ligation of purified DNA fragments

PCR of Alpha-link and link-A
Michał L. Marcin and Ewa

1. We've ran gradient PCR to obtain Alpha-link and link-A.
   Annealing temp 48°C - 55°C.
   Elongation time 1 - 2 h.
2. We have successfully amplified Alpha-link and link-A. Now its time for PCL (Polymerase Chain Ligation) to create fusions Alpha-A and Omega-A.