Team:Cambridge/Gibson/Protocol

From 2010.igem.org

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(Step 4: Gibson Assembly)
(Step 3: PCR)
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<td><b>Assembly Method</b>
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</td><td><b>Number of steps to ligate N pieces of DNA</b>
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<td><a href="http://partsregistry.org/Assembly:Standard_assembly" class="external text" title="http://partsregistry.org/Assembly:Standard_assembly" rel="nofollow">Standard Assembly</a>
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</td><td><a href="/Image:Steps-N.png" class="image" title="Image:Steps-N.png"><img alt="Image:Steps-N.png" src="/wiki/images/c/cc/Steps-N.png"  border="0" /></a>
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<td><a href="http://partsregistry.org/Assembly:Rolling_assembly" class="external text" title="http://partsregistry.org/Assembly:Rolling_assembly" rel="nofollow">Parallel Assembly</a>
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</td><td><a href="/Image:Steps-Log2.png" class="image" title="Image:Steps-Log2.png"><img alt="Image:Steps-Log2.png" src="/wiki/images/6/6f/Steps-Log2.png" width="138" height="21" border="0" /></a> (rounded up)
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<td><a href="http://2010.igem.org/Team:Cambridge/Gibson/Mechanism" class="external text" title="http://2010.igem.org/Team:Cambridge/Gibson/Mechanism" rel="nofollow">Gibson Assembly</a>
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The Annealing T<sub>m</sub> that should be used is the temperature of the main 20 or so bases of the primer (not including the flap), since the flap only begins to anneal after the first few cycles, by which point primer specificity is less of an issue.
The Annealing T<sub>m</sub> that should be used is the temperature of the main 20 or so bases of the primer (not including the flap), since the flap only begins to anneal after the first few cycles, by which point primer specificity is less of an issue.

Revision as of 19:32, 24 October 2010