Fuzzy sequencing gel bands

Robert Horton horton at molbio.cbs.umn.edu
Mon Aug 22 10:30:34 EST 1994


Bengt Oxelman (bengt.oxelman at systbot.gu.se) wrote:
: About a year ago, I obtained a sequence gel where fuzzy sequences occurred
: side-by-side with normal, sharp ones. The fuzziness was thus apparently
: reaction-specific. I don't think that it is a N-1 primer problem or due to
: sequence length polymorphism, because these problems cause double bands,
: not fuzzy bands. The fuzziness also tended to be more apparent at the
: bottom of the gel. I didn't bother too much at that time though, because
: the sequences could be well checked from the other strand.
: Now, I have got two gels with all sequences (different primers) fuzzy, and
: I am really frustrated, because I am in a real hurry.
: Does anyone have an idea on how I shall solve this problem? My methods
: include cycle sequncing (with initial cycle labling of 35S) and
: electrophoresis on a 5% acrylamide gel with a salt gradient.
: Any comments are greatly appreciated!
:

Sugar or glycerol in the sample can cause fuzzy bands at the bottom of the
gel...

 
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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