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Team:EPF-Lausanne/Protocols/T7-ext
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(Created page with "{{:Team:EPF-Lausanne/Templates/ProtocolHeader|title=T7 extension PCR}} This protocols consists in 3 steps: * Gene-specific PCR: amplifies the gene you want to put under the cont...") |
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* Extension PCR: adds the T7 promoter (or its variants) upstream the gene and overhangs for the Gibson assembly | * Extension PCR: adds the T7 promoter (or its variants) upstream the gene and overhangs for the Gibson assembly | ||
* Final PCR: amplifies the fragments extending the gibson overhangs | * Final PCR: amplifies the fragments extending the gibson overhangs | ||
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| + | == Gene specific-PCR == | ||
| + | |||
| + | Just a normal PCR to amplify the genes. | ||
| + | The primers should include the RBS and overhangs with the extension primers | ||
| + | |||
| + | == Extension PCR == | ||
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{{:Team:EPF-Lausanne/Templates/Footer}} | {{:Team:EPF-Lausanne/Templates/Footer}} | ||
Revision as of 12:54, 15 August 2011
T7 extension PCR
Back to protocols.This protocols consists in 3 steps:
- Gene-specific PCR: amplifies the gene you want to put under the control of the T7 promoter adding RBS upstream and overhangs for the next PCR
- Extension PCR: adds the T7 promoter (or its variants) upstream the gene and overhangs for the Gibson assembly
- Final PCR: amplifies the fragments extending the gibson overhangs
Gene specific-PCR
Just a normal PCR to amplify the genes. The primers should include the RBS and overhangs with the extension primers
Extension PCR
