Cosmid sequencing

Dave Fruhling Fruhlid at war.wyeth.com
Tue May 9 15:08:03 EST 2000


I am in pWe15 by stratagene with a 45kb insert.  I cannot make any of the primers work.  I have tried up to 100 cycles.  I have added 5-10% dmso or 5-10% gc melt.
my denaturaation is done 1x2min 98c and 100x 30sec 98c.  What am I not doing??? what can I do differently???
your help is greatly appreciated
David S. Fruhling
Deptment of Oncology
Wyeth Ayerst Research
914-732-2005
fruhlid at war.wyeth.com

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