hi there,
can somebody answer me just in a few words about the following
thoughts concerning (RNA) agarose electrophoresis:
1) why is the Maniatis et. al. buffer system for RNA gels based on
MOPS and not on TRIS ??? the pH of 7.0 should be covered by
TRIS as well ?
2) what is/are the main reason(s) for running the RNA in formaldehyde?
is it just unfolding it's secondary structure or additionally
prevent RNAase gegradation, or...?
3) somebody before pointed out to load EtBr together with the samples
onto the gel. we use EtBr mixed gel for DNA and this works fine.
since the EtBr is running in the opposite direction (to D/RNA), the
dye is removed from the nucleic acid the longer the gel runs.
when i started a gel today a big lane of EtBr was running off the
gel quickly. is this method still effective after long gels ??
it seems interesting to load EtBr in the lower buffer tank...
thnaks for any ideas
tom
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Tom Pasternak e-mail: ugn4501 at aberdeen.ac.uk
University of Aberdeen / SCOTLAND lab-tel: (224) 681818-53905
Ophthalmology / Immunology lab-fax: (224) 685158
ABERDEEN
AB9 2ZD
U.K.
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