Dr. G. Koraimann
koraimann at bkfug.kfunigraz.ac.at
Fri Aug 5 14:53:13 EST 1994
In article <315usd$69j at mserv1.dl.ac.uk> LMS <lee.smith at afrc.ac.uk> (Tel 0636 578411 X2598) writes:
>From: LMS <lee.smith at afrc.ac.uk> (Tel 0636 578411 X2598)
>Subject: RNA gels
>Date: 27 Jul 1994 16:35:41 +0100
>..I'm about do do my first RNA gel/Northern blot and I'm confused.
>..Why does everybody use formamide gels instead of glyoxal/DMSO
>..(which according to Maniatis give sharper bands)?
>..I would prefer not to use formamide as we don't have a fume
>..hood in the lab.
>..Any other ideas always welcome!
>..LEE SMITH, Institute for Animal Health,Compton
What I do when I run RNA gels:
In order to reduce toxic formaldehyde, I only use a sample buffer
containing this substance. I do not include formaldehyde in the gel. I tried
it several times and compared it with standard formaldehyde containing gels.
The result was that the resolution as well as the sharpness of the RNA bands
are comparable to the standard formaldehyde containing gels. The handling of
high percentage (>3%) agarose gels without formaldehyde is much better and
also there is no background from formaldehyde when visualizing the RNA with
Ethidiumbromide / UV irradiation.
You can find the details of the method (buffers, running conditions etc.) in
Mol Microbiol (1993) 9: 717-727. Originally the method was published in
Promega Notes, number 28, 1990
Institut für Mikrobiologie
A- 8010 Graz
AUSTRIA - EUROPE
Tel: (316) 380 5620
Fax: (316) 381548
e-mail: koraimann at bkfug.kfunigraz.ac.at
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