JROSENBAUM.CURRY@BIONET-20.BIO.NET (Alice Curry) (06/01/89)
Our lab is thinking about using non-radioactive labelling methods, such as biotinylated probes, for genomic southerns, northerns, and library screening. But we haven't found a lot of people with experience using them. I'd appreciate some advice on the subject-- what methods and kits have proved useful, which were not useful, how the costs compared to isotope labelling, was it worth making the switch? --Alice Curry -------
HUBERMAN@BIONET-20.BIO.NET (JOEL A. HUBERMAN) (06/03/89)
Alice,
We have a limited amount of experience with the "Genius" kit from
Boehringer-Mannheim. On good days it performs as advertised (0.1 pg
detected). On bad days, it's sensitivity may be as much as 10-fold lower.
Nylon membranes can be reprobed. For many purposes, it's as good or better
than 32-P labelling, and, of course, it's a lot safer. I haven't done a
detailed cost accounting, but, since the probes are stable and can be used
repeatedly, it's probably less expensive than 32-P.
I, too, would be interested in other investigators' experience in this
area.
Joel A. Huberman
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