Subcloning dirty PCR products

schupp at nauvax.ucc.nau.edu schupp at nauvax.ucc.nau.edu
Mon Aug 29 11:23:03 EST 1994


Sounds like incomplete digestion.  A few thoughts, sorry if you've already
had them.

        -purify the PCR product prior to restriction.
        -some enzymes are more sensitive to cutting at the end of a sequence
         than others.  New England Biolabs has some data on this in their
         catalog.
        -depending on the size of the fragment, you might be able to
         run the digested fragment on Synergel:agarose gel or a Metaphor
         gel against the uncut fragment to check the digestion.

Jim Schupp



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