Protocol- hybridization with oligonucleotides

txpljfg at uabcvsr.seas.ucla.edu txpljfg at uabcvsr.seas.ucla.edu
Tue Dec 3 00:18:00 EST 1996


We typically end-label oligonucleotides with 32P and hybridize in a 
tris-acetate high-salt buffer at 37-42 degrees (sometimes 55C depending 
on the particular oligo).  We wash in 1X SSC in 0.1% SDS at the same 
temperatures in the presence of sodium pyrophosphate, 1-2X Denhardts, and 
salmon sperm DNA.  I have a detailed protocol that I can email and/or 
post if you like.  Email me and I will do either.
-james-

Johanne Fleurent (johanne.fleurent at nrc.ca) wrote:
> Hello everybody,

> Do you have a good protocol for hybridization with oligonucleotides, it
> can be with radioactive or non-radioactive labels. 

> I also need a protocol to make a dot blot with yeast.


> Thank you.


> Johanne Fleurent
> Lallemand Inc.
> Montreal

--
==============================================================================
James F. George, Ph.D.              Transplantation Immunology Laboratory
Department of Surgery               *Please sign your organ donor card* 
University of Alabama at Birmingham
205-934-4261 voice
jgeorge at uab.edu
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