Seeking info re hybridising
hroychow at NMSU.EDU
Tue Nov 3 12:07:14 EST 1998
At 02:15 PM 11/3/98 +0100, Dr. S. Shapiro wrote:
> I'm a little new to the nucleic acid field, so forgive my asking
>what might be a simple-minded question.
> I am (in principle) able to generate a labelled NA probe, and I'd
>like to survey a population of bacteria to determine whether this population
>contains DNA complementary to this probe. I seem to vaguely remember
>reading that it is possible to apply whole-cell DNA to a special kind of
>filter, then add your probe, wash, and then see whether the probe sticks
>to the filter. The idea is that if the little probe sticks, then it is
>likely binding to a stand of complementary DNA.
> If anyone can suggest articles/books/reviews in which the above
>procedure is described _in detail_ with examples, kindly contact me
> toukie at zui.unizh.ch
>with the relevant information.
> Thanking all responders in advance, I remain
>toukie at zui.unizh.ch
What you are looking for is fondly known as "colony filter hybridization".
This is a very effective method for screening bacterial clones harboring
target sequences. You should be able to find the protocol in any mol. biol.
manual (eg. Sambrook, Fritz and Maniatis). If you do not have a ready access
to any, email me for a general protocol.
Dr. Hiranya Sankar Roychowdhury
Plant Genetic Engineering Lab.
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroychow at nmsu.edu
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