Screning E.coli transformants ...

Mike Prigge prigge at darkwing.uoregon.edu
Thu Feb 11 22:06:34 EST 1999


In article <bpmurray*STUFFER*-0902991653000001 at mac-daddy.ucsf.edu>,
bpmurray*STUFFER*@socrates.ucsf.edu (Bernard P. Murray, PhD) wrote:

> I'm in a lab of radiation-phobes/ecologically minded people
> and so the amount of stress from innumerable screenings is
> far less than from using a few microCi of 32P.
>      I've played with DIG/alkaline phosphatase Southerns
> in a previous lab with good success.  Does this work well
> for colony hybridisation or does anyone recommend any other
> non-rad methods for this?
>      Thanks,
>           Bernard

I used DIG/HRP to screen phage cDNA libraries and gridded BAC (colony)
libraries and was very pleased.  I can't comment on aDIG-AP for this kind
of application because I switched to HRP/luminol for all my
hybridizations.  I think I heard that AP detection can have problems if
you use nitrocellulose membranes.  If anyone knows if this is true (and
why), let me know as I'm curious.
Mike
-- 
Michael Prigge                          Phone: (541) 346-4256
Institute of Molecular Biology            Fax: (541) 346-5891
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