Non-radioactive detection systems

wetsel_r at wums.wustl.edu wetsel_r at wums.wustl.edu
Tue Dec 8 06:20:00 EST 1992


In a previous article, David_Nunn at qms1.life.uiuc.edu (David Nunn) wrote:
>   I'm sure that this same thread has been on the net before but forgive
>me. 
>   We are thinking of switching to non-radioactive Southern hybridization.I
>would like some feedback on which systems you have found to be the easiest
>to use, most cost effective, etc..  We would be using these methods
>primarily for clone screening and prokaryotic chromosomal DNA hyb's, so
>extreme sensitivity is not required.  If I get a few responses I will
>repost them to the net.  Thanks
------
David:

We have tried Amersham's ECL 3'-oligo labeling kit as an alternative to 
radiation and it has worked very the couple of times we've used it.  It was 
one of the few "kits" that managed to work the first time when used by the 
book.   We weren't looking for anything rare, our Southern was an oligo 
that hybridized to digested fragments of a cosmid.  Unlike radiation, one 
can't follow the probe so until you go through the developing steps you 
don't know if you've inadvertantly washed your probe off - 

Soon, when some $$$ come in, we'll be trying the ECL random prime kit for 
Southerns and Northerns.  I don't know about anyone else's institution, but 
here at Wash-U we're looking at about $500 everytime a 5 gallon drum of 
liquid radiation waste leaves the lab...  if for nothing else, that is 
gentle "push" to consider non-radioactive alternatives...

Best of luck,

David
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+  David L. Haviland, Ph.D.	     Internet:"haviland at kids.wustl.edu"   +
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