PCR product cloning problems

Bill Rogers brogers at noguchi.mimcom.net
Wed Aug 14 09:55:23 EST 2002


I recently cloned and sequenced a batch of PCR products and found that
many of the products were missing 1-10 bases from the ends. To me this
could be either

1. lousy primers (and I'm checking that now)
2. exonuclease activity during the PCR reaction (I used the
Boehringer-Mannheim core PCR kit, Taq polymerase)
3. exonuclease activity during cloning (using the PCR Script kit)

Has anyone here had similar problems or have any suggestions to avoid
this?

Thanks for any help you can offer.

Bill



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