We are having a problem cycle sequencing microsatellite clones and are
wondering if any of you out there have solutions or ideas. The problem
is not unique, as we have spoken with other researchers who have
observed similar phenomena.
When sequencing through clones, the reactions often die immediately
following the microsatellite, that is, they just degenerate into very
poor sequence. It doesn't happen with all clones, and it doesn't even
consistently happen with the same clone. It also appears to have
nothing to do with the length of the microsatellite, as we have obtained
beautiful sequence through dinucleotide repeats as long as 140 bp. We
are sequencing directly from the plasmid.
Thanks for any help you can offer.
Stacia Engel
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Stacia R. Engel, Ph.D.
Research Microbiology and Genetics
E. & J. Gallo Winery
P.O. Box 1130
Modesto, CA 95353-1130
(209) 341-8166 phone
(209) 341-8167 fax
stacia.engel at ejgallo.com
