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Team:Warsaw/Calendar-Main/25 June 2008
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| - | + | <h3>Preparation of pMPMT5-AID+AIDT7 construct</h3> | |
| - | < | + | <p><ol> |
| - | + | <li> Digest of isolated PCR product (from previous day) and pMPMT5-AID with HindIII and XbaI, dephosphorylation of pMPMT5-AID with CIAP </li> | |
| - | + | <li> Clean-up of digested PCR product. </li> | |
| - | + | <li> Gel electrophoresis of digested pMPMT5-AID and gel-out of proper band (4800 bp)</li> | |
| - | + | <li> Overnight ligation of purified DNA fragments</li></ol> | |
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| - | We've ran gradient PCR to obtain Alpha-link and link-A. Annealing temp 48°C - 55°C. | + | <h3>PCR of Alpha-link and link-A<br> |
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| + | Michał L. Marcin and Ewa</h3> | ||
| + | <p>We've ran gradient PCR to obtain Alpha-link and link-A. <br>Annealing temp 48°C - 55°C.<br> | ||
| + | Elongation time 1 - 2 h.</p> | ||
| - | 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. | + | <p>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. </p> |
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Revision as of 23:16, 1 October 2008
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Preparation of pMPMT5-AID+AIDT7 construct
PCR of Alpha-link and link-A
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