Isolation of Total RNA from just about anything

dkim at unm.edu dkim at unm.edu
Thu Jun 30 12:22:12 EST 1994


(Post about running RNA on gels omitted)

First: Denaturation of secondary structure in RNA for gels is also necessary
if you want to get an accurate estimate of molecular weight, since folded RNA
will run faster than the relaxed single strand.

Second: If your only concern is seeing the RNA, you might want to try just 
quickly running a bit on a regular TBE-agarose gel (maybe 1 %). Then stain
with ethidium bromide. The rRNA bands will show up (although not at the
"right" places) and the ethidium bromide fluorescence will be enhanced by
intercalation into dsRNA. (Fluorescence is not so good on ss strands). In 
addition, you won't get the awful background that is typical of formaldehyde/
formamide gels.

Third:  You may consider using a different loading buffer, in case the Sigma
stuff is not quite nuclease free.

Daniel Kim



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