polyA problem with ESTs

Olin Anderson oandersn at pw.usda.gov
Wed Dec 19 15:33:08 EST 2001


We have a large EST project and are having trouble with the
the 3' sequencing - reading through the polyAs.  More than
30 A's and the sequence is short, longer than 50 and we get
no good sequence.  I've heard some sites (but not all) have
had success with the oligodT anchored primers.  Could anyone
share the conditions/primers/etc they use to successfully get
long reads past cDNA polyA stretches?  Thanks.

Olin

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