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Team:Warsaw/Calendar-Main/26 June 2008
From 2008.igem.org
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| - | <h3>Preparation of pMPMT5-AID+AIDT7 construct </h3> | + | <h3>Preparation of pMPMT5-AID+AIDT7 construct <br>Michał K.</h3> |
| - | <ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of E. coli <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> from previous day </li> | + | <ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> from previous day </li> |
| - | <li>Plating transformants on LB + tetracycline</li> | + | <li>Plating transformants on LB + tetracycline</li></ol> |
| - | <h3> Polymerase Chain Ligation (PCL) </h3> | + | <h3> Polymerase Chain Ligation (PCL) <br>Michał L., Ewa, Marcin</h3> |
<table id="result"> | <table id="result"> | ||
Revision as of 15:33, 9 October 2008
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Preparation of pMPMT5-AID+AIDT7 construct
|
| Product | Templates | Primers | Product length |
|---|---|---|---|
| Alpha-A fusion | Alpha-linker + linker-A | AlphaL+SacI + AP+NotI | 1500 bp |
| Omega-A fusion | Omega-linker + linker-A | OmegaL+SacI + AP+NotI | 1440 bp |
PCL program for fusion A with alpha and omega
| Temperature | Time | |
|---|---|---|
| 94°C | 4:00 | |
| 94°C | 0:30 | 28 cycles |
| 48°C-62°C | 0:45 | |
| 72°C | 2:30 | |
| 72°C | 10:00 | |
| 4°C | infinite |
