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Old 08-14-2013, 02:33 PM   #5
Catherine_Burke
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Location: Sydney, Australia

Join Date: Oct 2012
Posts: 6
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Thanks for all your replies.

I did see the Lahr and Katz (2009) paper, which is what got me worried about phusion! They do show very little recombination with small amounts of starting template, but I'm unlikely to have less than 10^5 copies in an enrichment PCR. Above this template concentration, they see ~50% chimera formation with 39 cycles (they didn't test less than this).

Another paper indicates that recombination will be more of a problem with more complex templates (like microbial communities!)
http://www.ncbi.nlm.nih.gov/pubmed/22278883

And here is the link to the paper which suggests proofreading polymerases cause more chimeras than regular taq (although I don't have access so have only read the abstract!)
http://link.springer.com/article/10....275-012-2642-z

From speaking to a few other people, it seems like most use a regular old Taq, so I'll probably try that for next time...
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