Rna gel

MELANSOND melansond at am.abru.cg.com
Tue Dec 3 09:24:49 EST 1996


***Can someone give a good recipe for a formaldehyde/agarose gel for rna. I
***see all kinds of different concentrations etc.. Does someone have one,
***that works consistantly. ALso I put etbr in my gels, is it better to
***stain later or in the gel. I know it decreases signal intensity, but I
***like seeing my rna. THanks
***
***-- 
***William Wu
***PhD student
***McGill University
***Dept of Experimental Surgery
***Shriner's Hospital For Crippled Children
***1529 Cedar Ave. 
***Mtl. PQ. Canada
***H3G 1A6
***http://www.geocities.com/yosemite/1156/index.html
Hi
	What consistantly works for me is:

	1.2 g agarose
	86.6 ml water
heat to dissolve agarose, cool to 65 degrees
	Add 10 ml 10X MOPS
	     3 ml 37% Formaldehyde
Mix gently but thouroughly, pours 100 ml gel.


	then the loading dye should contain EtBr, I don't add to the gel or
stain, but directly to the RNA sample for the best intercalation and
visualization

	Loading Dye
	500 microlitres formamide/0.1% bromophenol blue
	170 microlitres formaldehyde
	100 microlitres 10X MOPS
	  4 microlitres 10mg/ml EtBr
	225 microlitres sterile water

Add directly to RNA pellet and suspend, heat at 65 for 15 min, then flash chill
on ice and load gel.

Good luck




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