Team:Cambridge/Protocols/PCRcolony

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Colony PCR:

Used to amplify DNA directly from cell culture, the reaction is the same as for normal PCR but with modified reaction composition and cycle settings, here shown for a 50 µl reaction using Taq polymerase.

ReagentVolume (µl)Final Concentration
Water35.7
10 mM dNTPs1200 µM
10 x NH4 buffer51x
Forward Primer2.50.5 µM
Reverse Primer2.50.5 µM
Template Cells1.3 (from liquid culture or picked colony)
Taq polymerase 5u/µl10.1 u/ µl

Please refer to the standard PCR protocol for the remainder of this protocol.


Notes on colony PCR

  • This technique should only really be used for crude PCR assays, such as diagnostic PCR. If high quality DNA is desired, Miniprep followed by standard PCR should be used.



Risk Assessment


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