RNA electrophoresis!!

Han huangz at hotmail.com
Fri Oct 31 17:55:08 EST 1997


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)

10xMOPS: 
        MOPS  41.2 g
  	Sodium acetate 6.57 g
  	EDTA,sodium salt  3.7 g
  	water to 1 litre(pH to 7.0)

1xMOPS gel running buffer: add EB to final concentration 1ug/100ml

1% formaldehyde agarose gel:
	37% formaldehyde	8.5 ml
	RNase free water	37.5 ml
	10xMOPS			5 ml
	Agarose			0.5 ml
	EB			25 ug

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

Thanks in advance.

HZ
Gut hormone Lab
K.U.Leuven



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