PCR cleanup before digest

newsnet customer customer at newsnet.com
Thu Mar 30 03:26:37 EST 2006


How does proof-reading enzymes affect downstream cloning?

ST

  More importantly you would want to get rid of the polymerases, especially if they have proofreading activity (ie in case you want to clone it downstream). 
  If you just want to digest without downstream cloning, I see no problem for most enzymes to do it directly from the PCR reaction.

  AS
    "newsnet customer" <customer at newsnet.com> 
    Hi,

    Been told to do a PCR cleanup before doing a restriction digest because the salt (MgCl2) in the PCR mix can affect cleavage efficiency. Anyone like to comment?


    ST



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