need HELP with non-radioactive Northern hybs

Tracy Aquilla aquilla at salus.med.uvm.edu
Fri Mar 11 16:57:18 EST 1994


Greetings gene jockeys,
  I am interested in using non-radioactive labeling for quantitative
Northern blot analysis, and I need help deciding which system to use. Any
comments from experienced users of such products would be sincerely
appreciated. I need high sensitivity for this assay, and I prefer to use
oligonucleotide (ssDNA) probes.
  I think the LightSmith (Promega) and Aces (BRL) type of approach seems to
be the best idea I have seen so far, but I am reluctant to put an
AP-conjugated oligonucleotide through the rigors of a stringent
hybridization protocol, for fear that it will inactivate the enzyme. Also,
what about HPR-conjugated oligos? Has anyone used these or similar products
with success?
  Please respond by email if you can help. Thanks in advance.
Tracy Aquilla, Post-doctoral Research Associate
Department of Molecular Physiology and Biophysics
University of Vermont, College of Medicine
aquilla at salus.med.uvm.edu



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