getting through polyG regions

William J.Buikema wjb1 at midway.uchicago.edu
Thu Apr 12 09:18:45 EST 2001


Hi,
    I have a couple of users that have inserts containing polyG
stretches of 12 to 18 bases.  There seems to be a strong cutoff in
signal after 12-13 consecutive G's, and I have not had any success
getting through longer stretches.  Should this be treated simply as high
GC templates (ie. try DMSO, GC kit, 98C denaturation, betaine, etc), or
is it a special case because they are all G's?  Anyone else out there
solve this problem?
    I'm using BD 2.0 on 377XL's, and have tried 98C denaturation, and
double reaction mix to no avail.

TIA,

Bill Buikema
w-buikema at uchicago.edu

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