Path: utzoo!utgpu!watmath!iuvax!bionet!bionet-20.bio.net!BMCCARTHY.LUDWIG
From: BMCCARTHY.LUDWIG@BIONET-20.BIO.NET (Erwin Ludwig)
Newsgroups: bionet.molbio.methds-reagnts
Subject: ligation of pcr products
Message-ID: <12486092662.25.BMCCARTHY.LUDWIG@BIONET-20.BIO.NET>
Date: 14 Apr 89 15:55:30 GMT
Sender: daemon@NET.BIO.NET
Lines: 4


  Tacking on a restriction site on your pcr oligos helps alot, such as using a 24mr with a Hind III or Eco RI site in it.It shouldn't make a difference if the 5 prime end is floating in the breeze.  Another more obvious solution is kinasing the blunt end fragment, since pcr oligos do not have 5 prime phosphates they will not insert well.  Add a pinch of 32P gamma label when end labeling and run it on a gel prior to ligation, the 32p will not interfere with ligation.  It wouldn't hurt to phosphatase your ve
ctor to.
Erwin
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