Team:Cambridge/Gibson/Protocol

From 2010.igem.org

(Difference between revisions)
(Step 3: PCR)
(Step 3: PCR)
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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.
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The Polymerase mixture we used was[http://www.finnzymes.com/pcr/phusion_high_fidelity_pcr_mastermix.html 2x Phusion MasterMix].
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The Polymerase mixture we used was [http://www.finnzymes.com/pcr/phusion_high_fidelity_pcr_mastermix.html 2x Phusion MasterMix].
==Step 4: Gibson Assembly==
==Step 4: Gibson Assembly==

Revision as of 20:39, 23 October 2010