Team:Warsaw/Calendar-Main/25 June 2008

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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

Michał L. Marcin and Ewa

PCR of Alpha-link and link-A

We've ran gradient PCR to obtain Alpha-link and link-A. Annealing temp 48°C - 55°C. Annealing time 1:00 - 2:00. 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.