Path: utzoo!utgpu!watserv1!watmath!uunet!shelby!snorkelwacker!bionet!cluster.sussex.ac.uk!BAFM1
From: BAFM1@cluster.sussex.ac.uk
Newsgroups: bionet.molbio.methds-reagnts
Subject: re:pcr sequencing
Message-ID: <9010012312.AA05694@genbank.bio.net>
Date: 1 Oct 90 11:08:00 GMT
Sender: daemon@genbank.bio.net
Lines: 9

In reply to Doug Prashers query RE problems with sequencing PCR products,
it is important (at least with klenow) to NOT have the buffer present
(ie. TM) during the annealing step. It is added at the sequencing reaction
stage. The presence of ss binding protein also helps. I must admit that I
prefer to reclone into M13, but direct sequencing is useful if you just want
to check a short region.
Hope this helps,
Jim Brannigan
BAFM1@UK.AC.SUSSEX.CLUSTER