sequenase kit

Brain Foley brianf at
Fri Jun 19 13:09:11 EST 1992

The problem with ds-DNA templates is not always with the purity of the
template DNA.  The denaturation/anealing steps can have a big impact
on the quality of the sequence.  There are numerous ways to deanture
and aneal the primer.

The best way to trouble-shoot is to get a plasmid and primer that is
known to work in someone elses lab, and run that side-by-side with
your plasmid and primer.  

Remember that Murphy's law says that if anything can go wrong it will.
In sequencing there are MANY things that can go wrong.  It is truly
amazing that it usually works so well in experienced hands.  Getting
the experience is not a waste of time, if you never experience the
various troubles, you never learn how to trouble-shoot and you
never learn what the important parameters are.
*  Brian Foley               *     If we knew what we were doing   *
*  Molecular Genetics Dept.  *     it wouldn't be called research  *
*  University of Vermont     *                                     *

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