ssDNA by PCR-Asymm or exonuclease?

Mike Morris mike at medsun.unige.ch
Wed Feb 22 06:52:20 EST 1995


In article 200295143555 at 132.236.8.54, gsr1 at cornell.edu (Geoffrey S. Rule) writes:
>>Hello folks,
>>	Was just wondering if  anyone has experience, or comments to offer,  with
>>either of the methods for producing ssDNA, ie, by use of exonucleases or by
>>asymmetric synthesis. 
>>Geneva, NY 14456 <<-- Coincidence! :)

  We used to do asymmetric PCRs, with a twist: you do a first, normal
PCR; then use 1 ul of that as template for a second PCR, to which
you add only one oligo. Thirty cycles, of course. Doing this, we used 
to get good sequence with T7 pol on about 50% of samples - and no
sequence on the rest (we have now switched to cyclic sequencing for
PCR products, using Sequitherm - wonderful!).

  Mike Morris

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