Re: Sequencing PCR Products onthe abi377XL

HOSSEINI@gmm.gen.emory.edu
Fri, 24 Jul 1998 8:59:53 EST5EDT

From: "robert obrien" <rfob@email.msn.com>
Subject: Sequencing PCR Products onthe abi377XL
Date: Thu, 23 Jul 1998 22:20:39 -0400
To: Recipients of ABRF List <abrf@aecom.yu.edu>

I have recently begun sequencing 200bp pcr products on the 377XL and have
been noticing the following abnormalities: Peaks have a flat top appearance
to them and there are peaks under larger ones. Is it customary to get a lot
of N calls? What% is acceptable to get?

Any Suggestions will be greatly appreciated

Thank You

Robert O'Brien

Robert:

What technique do you use to get rid of excess dye after cycling? One possible explanation
for high peaks with flat top and having real peaks under larger one is that excess dye are
there. We always use CentriSep columns to clean the cycling products and it works perfect.

Hope this helps

Seyed

Seyed H. Hosseini, Ph.D.
1462 Clifton Rd. Rm#416
Center for Molecular Medicine
Emory University
Atlanta, GA 30322
Tel # 404-727-1353
Fax # 404-727-8367
hosseini@gmm.gen.emory.edu