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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