RNA electrophoresis!!

glover martin gwmartin at fas.harvard.edu
Fri Oct 31 13:46:36 EST 1997


Han <huangz at hotmail.com> wrote:
> Dear netters,

> I encountered a problem in RNA formaldehyde electrophoresis: I added EB 
> in my formaldehyde agarose gel(final concentration 50ug/100ml) and run 
> gel for 2 hours. Amazingly, when I visualize gel under UV lamp I found 
> very high staining background which evenly distributed and composed 
> almost 2/3 area of gel starting from the margin near the wells. This high 
> flurescent staining is so strong that overhided my RNA bands. I would be 
> very appreciate any suggestion about the possible reasons. My RNA 
> formaldehyde electrophoresis buffers are as following: (please save me 
> out of the nightmare)


> RNA loading buffer:
> 	RNA (10-20 ug)		5 ul
> 	10xMOPS			1 ul
> 	37% formaldehyde	3.5 ul
> 	formamide		10 ul

I put ethidium bromide in the loading buffer. Am I the only one?

Trei



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