Path: utzoo!utgpu!water!watmath!uunet!ig!bionet-20.bio.net!EROSENTHAL
From: EROSENTHAL@BIONET-20.BIO.NET (Eric Rosenthal)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Uniform gels for reading long sequences
Message-ID: <12412284971.17.EROSENTHAL@BIONET-20.ARPA>
Date: 7 Jul 88 02:37:44 GMT
Sender: daemon@presto.ig.com
Lines: 20


	We have been sequencing, using sequenase, and running the reactions
on a 8% gradient gel.  This gives us 250 to 300 bases worth of sequence.
In many cases we have then run a 6% uniform gel to try and extend the
amount of sequence information from the same set of reactions, but we
have had very mixed results with this.  The gels are often quite blurry
and we get very little additional sequence information from them.  Does
anyone have any suggestions for how to improve the quality of these
extended gels.
	A related question-- We tryed wedge spacers with these long gels
and the speration of bands was quite good, but we had terrible problems
with handling the gels.  In particular, the gels never dryed completely
on the gel dryer.  Even when we thought they were dry they would stick to
the X-Ray film and the autoradiographs were often covered with little spots.
A number of other people have told me that they have had the same problems.
Does anyone have any tips?
	Thanks.

						Eric Rosenthal
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