nospam_jweiner1 at ix.urz.uni-heidelberg.de
Tue Sep 1 08:43:34 EST 1998
[ defaultuser at domain.com ]
> I'm starting to work with RNA-Gels. But I haven't got any idea if it's
> possible to SEE mRNA on an agarose-ethidiume-bromide-gel. I see two
> spots and I'm sure that is the rRNA. I induce a typical genexpression,
> but I'm not sure if it's possible to see some difference between control
> and stimulated cells.
1) maybe you could try to fit your signature in four lines?
2) of course you can see RNA on agarose gels stained with EtBR. The
only problem is to get the RNA into denaturated state. I usually achieve
this by heating the sample to 95 deg C for 2 minutes diluted 1:1 with a
loading solution containing 7-8M Urea and the dyes (xylene cyanol and
bromophenol blue). You can also try to make formaldehyde gels, but they are
a mess and you don't get much better results with them. If the RNA is not
denaturated, you will see a smear only.
3) if you are not sure, whether you see RNA or not, you can treat
your gel with RNAse - CAUTION!!! Anything contaminated with Rnase must not
get near your bench space. As a negative control you can treat your gel
with DNAse to see whether the bands you see are DNA.
Hope that helps,
Scziastie dla wsiech, darom i pust' nikto nie ujdiot obiýennyj
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