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
===============================================================================
More information about the Methods
mailing list