Team:Cambridge/Gibson/Protocol

From 2010.igem.org

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(Step 4: Gibson Assembly)
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==Step 1: Design Primers==
==Step 1: Design Primers==
{{:Team:Cambridge/Templates/RightImage|image=Cambridge-oligoface.jpg|caption=Designing Oligos Old-School}}
{{:Team:Cambridge/Templates/RightImage|image=Cambridge-oligoface.jpg|caption=Designing Oligos Old-School}}
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{{:Team:Cambridge/Templates/RightImage|image=Gibthon.png|caption=New and improved Gibthon oligo design}}
*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==
==Step 2: Order Primers==
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{{:Team:Cambridge/Templates/RightImage|image=Gibthon.png|caption=New and improved Gibthon oligo design}}
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*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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{{:Team:Cambridge/Templates/RightImage|image=Phusion.jpg|caption=Phusion Polymerase}}
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==Step 3: PCR ==
==Step 3: PCR ==
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{{:Team:Cambridge/Templates/RightImage|image=Phusion.jpg|caption=Phusion Polymerase}}
PCR is a dark art, but we have found that these general principles have served us well over the summer.
PCR is a dark art, but we have found that these general principles have served us well over the summer.

Revision as of 20:41, 23 October 2010