Team:Cambridge/Gibson/Protocol

From 2010.igem.org

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(Step 4) Gibson Assembly)
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The formal paper in nature describing Gibson Assembly can be found [http://www.nature.com/nmeth/journal/v6/n5/full/nmeth.1318.html here].
The formal paper in nature describing Gibson Assembly can be found [http://www.nature.com/nmeth/journal/v6/n5/full/nmeth.1318.html here].
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==Step 1) Design Primers==
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==Step 1: Design Primers==
*If you wish to ligate two pieces of DNA using Gibson they must be altered so as to have 40bp of overlap at the point of ligation.
*If you wish to ligate two pieces of DNA using Gibson they must be altered so as to have 40bp of overlap at the point of ligation.
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==Step 2) Order Primers==
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==Step 2: Order Primers==
*This step can take a while, so Gibson Assembly requires some planning ahead
*This step can take a while, so Gibson Assembly requires some planning ahead
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==Step 3) PCR ==
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==Step 3: PCR ==
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==Step 4) Gibson Assembly==
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==Step 4: Gibson Assembly==
*Prepare [[Team:Cambridge/Gibson/MasterMix | Master Mix]]
*Prepare [[Team:Cambridge/Gibson/MasterMix | Master Mix]]

Revision as of 13:16, 21 October 2010