RNA gels - staining and denaturing

Oleh Kovalchuke oleh at post.its.mcw.edu
Sun Aug 6 13:45:43 EST 1995

Andrea Moriondo (amoriond at galactica.it) wrote:
: amarion at moose.uvm.edu (Amy Marion) wrote in article 
: >
: >What is the proper procedure for staining RNA gels (either glyoxal or 
: >formamide) with ethidium bromide? 
: > 
The proper procedure can be found in Sambrook-Maniatis.
Here are two improper ones:
= load samples denatured with formamide or urea (both work fine) on regular 
DNA gel, i.e.
agarose/TAE/EtBr - see S.-M. manual for recipe.  Separation of rRNA
bands is as good as with formamide gels, the bands are _a bit_ fuzzier.
Sensitivity is about 50 ng.  I use this method for hassle-free detection of
rRNA as well as for cleaning of rRNA after labeling.
= Add 1-10 ug of EtBr to sample after heating.  Separate on formamide gel.
Sensitivity is about 1 ug rRNA.

Oleh Kovalchuke
at home

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