rphjr at nwu.edu
rphjr at nwu.edu
Fri Apr 1 23:29:16 EST 1994
In article <1994Mar31.162243.1 at molbiol.ox.ac.uk>, polk at molbiol.ox.ac.uk
> In article <1994Mar31.153500.1 at molbiol.ox.ac.uk>, polk at molbiol.ox.ac.uk writes:
> > In article <1994Mar31.123756.460 at chmeds.ac.nz>, mkennedy at chmeds.ac.nz (Martin Kennedy) writes:
> >> In article <2n2ha1$39c at quartz.ucs.ualberta.ca>, asimmond at gpu.srv.ualberta.ca (Andrew Simmonds) writes:
> >>> I am sure this has been covered before....
> >>> I am having trouble getting TIGHT bands with Northern blots from
> >>> formaldehyde RNA gels... When I cut off the ladder from the gel and look
> >>> at it sideways the bands are "tilted" i.e. ( / ) and I am sure this is a
> >>> problem when I transfer the RNA to a membrane causing the wide bands I am
> >>> seeing. I put 2.2M formaldehyde in the gel and use a standards MOPS
> >>> buffer...
> >>> Might anybody suggest a better way?
> >>> Andrew Simmonds
To Northern runners,
Formaldehyde is a wonderful chemical to use to keep RNA denatured, but it
is a known carcinogen and is rather toxic. Why use it if you don't have
to. You don't have to use it in RNA Northerns. You can use a glyoxylation
protocol to denature Your RNA and keep it denatured. I too was a sceptic,
being a formaldehyde man for years, until I tried it. I've never had
better Northerns and I don't have to work with a well known carcinogen! So
my suggestion to you is that if you feel formaldehyde is at the root of
your problem(sideways bands and inefficeint transfer), eliminate it. If
you want the ever so easy glyoxylation protocol, e-mail me. I am happy to
rphjr at merle.acns.nwu.edu.
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