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 -------