Path: utzoo!attcan!uunet!husc6!linus!philabs!aecom!diaz
From: diaz@aecom.YU.EDU (Dizzy Dan)
Newsgroups: sci.bio
Subject: Obtaining single-stranded linear DNA
Message-ID: <1792@aecom.YU.EDU>
Date: 11 May 88 03:49:37 GMT
Organization: Graduate School of Hard Knocks
Lines: 16

Here's this week's biochemical puzzler (you all did well on the
antisense RNA trick question):

I am interested in obtaining tritiated (or otherwise labeled)
single-stranded linear DNA of uniform length (has to be at least 1knt
long) for exonuclease assays.

M13 seems ideal since the cells will spit out labeled ssDNA (circular)
if we cook them with label.  How do we make the DNA linear?  Do we shear
it, or cut it with a restriction endonuclease with ss activity
(expensive!) or what? The DNA does not have to be linearized in the same
place, but it must be linear, and cut once, or at most a few times.

-- 
      dn/dx      Dept Molecular Biology   diaz@aecom.yu.edu
     Dizzy Dan   Al Einstein's Med School  Big Bad Bronx, NY