Team:EPF-Lausanne/Tools/Gibson assembly

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We successfully used Gibson assembly for our plasmid assemblies, allowing us to put different genes in our plasmid backbones. You can find our protocol in our Notebook, under "Protocols" section.
We successfully used Gibson assembly for our plasmid assemblies, allowing us to put different genes in our plasmid backbones. You can find our protocol in our Notebook, under "Protocols" section.
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== Requirements ==
 
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In order to perform Gibson assembly on two DNA sequences, these sequences need to overlap by at least 20bp at the edge. You can easily add these overlaps by extension PCR
 
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== Disadvantages ==
== Disadvantages ==
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* Primers are more complicated to design and longer - they are more expensive and take longer to synthesize
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* Primers are more complicated to design and longer - they are more expensive and take longer to be produced
* The reaction mastermix is expensive - due to large amounts of Taq ligase needed
* The reaction mastermix is expensive - due to large amounts of Taq ligase needed
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* A lot of negative resluts, du to the tendency of the backbones to close up themselves
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* A lot of negative results, due to the tendency of the plasmid backbone to close up on itself

Latest revision as of 21:41, 18 September 2011