genomic southerns

Hiranya S. Roychowdhury hroychow at NMSU.EDU
Fri Aug 20 10:17:01 EST 1999


At 11:40 AM 8/20/99 GMT, soenke behrends wrote:


>
>We would be very happy about any comment (or 
>magic trick) or good working protocol.
>
>Thanks a lot for your time and help
>
>Soenke Behrends
>

Frank has outlined the parameters very nicely. I have had quite a lot of
trouble with plant genomic Southern. One of the things that a colleague, in
another lab here, does is that she digests a 'huge' amount of the DNA over
two days to ensure complete digestion of 'more than sufficient' target and
THEN determines the concentration of the DNA spectrophotometrically  (Uncut
whole genomic DNA from plants is hard to quantitate). At least 10ug of the
*digested DNA* are loaded per lane. The hybridization protocols are too
different from lab to lab, so one should optimize within his/her
environment. I have switched to Church's for simplicity.


Dr. Hiranya Sankar Roychowdhury
GENE LAB/ EPPWS
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroychow at nmsu.edu




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