PCR product stability

Robert Horton horton at molbio.cbs.umn.edu
Mon Nov 14 19:55:12 EST 1994


Bengt Oxelman (bengt.oxelman at systbot.gu.se) wrote:
: Today I checked a couple of PCR products that were gel/Qiaquick purified
: about two months ago. We have recently detected pH problems with our
: milli-Q water so I wanted to check that the fragments were still there.
: They were, apart from a few acid solutions. But, all sample also contained
: a second fragment of half the size of the orignal fragment (two different,
: 1.1 and 1.6 kb)!!!!!!! ?????????
: 
: Does anyone have an idea about what is going on? Can I safely sequence the
: DNA (produced lovely sequences two months ago)?
:

Were they stored in plain water? DNA, particularly short pieces like PCR
products, can denature at low temps in ddH2O - maybe you were seeing
the two individual single strands? If this is the case, they should go away
if you add buffer, heat, and slowly anneal.... Jus' a thought.

 
Bob Horton (Ph.D.!)   /\ "Crash programs fail because of the theory that
U. of Minnesota, CBS  || with nine women pregnant you get a baby a month" 
1479 Gortner Ave.    /||\   -Werner von Braun.  Disclaimer:"Bob who?"
St. Paul, MN 55108    ^^   horton at molbio.cbs.umn.edu/(612) 624-3790



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