RNA gels - staining and denaturing

[dinakar bhattramakki]

Amy Marion (amarion at moose.uvm.edu) wrote:
: Dear netters;

: 	I need some advice on RNA gels.  I have made 3 attempts to run
: total RNA through glyoxal/DMSO gels and one attempt with a formamide gel.
: In all cases I have been unable to see the RNA after ethidium bromide
: staining.  Even the RNA markers do not show up.

: Should RNA be stained with ethidium bromide at all?  (These gels will be
: used for Northerns.)
: What is the proper procedure for staining RNA gels (either glyoxal or
: formamide) with ethidium bromide?

: 	I have run the RNA through 0.1% SDS, 1.2% agarose gels.  After
: staining with EtBr all the RNA (including the RNA markers) are visible.
: Will the 0.1% SDS denature the RNA?  If yes, can the RNA from this gel be 
: transfered to nylon for a Northern?

: 	Thanks for your help.

: Amy Marion
: marion at smtplink.ipfw.indiana.edu


Just use 1 ul of 0.4 ug/ul for 50 ul RNA solution (containing 10 ug 
RNA). This should difinitely stain very well two rRNA bands and your 
marker. 
-Dinakar
--
Dinakar Bhattramakki/Ph.:(402) 472-6243/email:ddh at unlinfo.unl.edu