Northern Blotting despair!
hroychow at NMSU.EDU
Thu Aug 14 15:52:40 EST 1997
At 09:08 AM 8/14/97 -0700, a.mccormick wrote:
>Does anyone have a reliable Northern Blotting method that will work
>using total or messenger RNA and probing for low abundance sequences??
>As you can tell we are having rather a terrible time with northern
>ad.mccormick at ulst.ac.uk
If you want help with anything, you should provide details of your woe.
'Northern blotting' does not tell us anything. For example, in this lab,
students refer to a simple RNA gel as "northern". By extension, some
students (and some investigators too) have been heard referring to protein
PAGE as "western". I used to rectify them, but have given up now. Anyway, if
you want a protocol for "northern" analysis, just grab either the "Red" or
the "Blue" book. The methods are not that complicated. As for making sure if
the "abundance" of the transcript is be the limiting factor, just try
hybridizing the "blot" with a probe for a housekeeping species (eg. actin).
This will also make sure that the "transfer" worked (alternatively, simply
stain a (portion of) blot with metylene blue, or, if EthBr has been used in
the gel or in the sample loading buffer, simply visualize the RNA under UV
on the membrane.
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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