Karyn,
We use spin columns from Bio-Rad(Cat#732-6226)for PCR product sequencing.
We have no problems with dye blobs or loss of signal.
Nathan J. Bivens
DNA Core Facility
M442 Medical Science Building
University of Missouri
Columbia, MO 65212
-----Original Message-----
From: Karyn T O'neil [mailto:Karyn.T.ONeil@dupontpharma.com]
Sent: Wednesday, December 20, 2000 9:37 AM
To: Recipients of ABRF List
Subject: Sequencing PCR products
Does anyone have a recommendation for sequencing PCR products? We find
that if the sequencing reactions are precipitated with isopropanol we have
a significant problem with dye blobs. When we use columns to clean up the
unincorporated dyes, we end up losing a significant amount of the signal.
We have only tried columns from EDGE and I am wondering if anyone has
experience with a column where you do not lose so much DNA but the dye
blobs are effectively removed.
Karyn O'Neil
Sr. Research Scientist
DuPont Pharma. Co.
Applied Biotechnology Dept.
P.O. Box 80336
Wilmington, DE 19880-0366
phone: 302-695-1227
fax: 302-695-9420
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