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Denaturing Agarose gel for RNA - (Apr/19/2006 )

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huh.gif Hi, i want to run a RNA gel to check the integrity of the RNA, is necesary to do it with a denaturing agarose gel, or is it possible to run a simple agarose gel????

MCR

-MCR-

If you run normal agarose gel you will be able to "see" your RNA but you won't get the information about the quality of the RNA sample...
All the best,

Helisa

-helisa-

but if you run on E-gel (like 2%), then you could also see the integrity of your RNA sample.
good luck.

-six-

I use 1.2% E-gels (invitrogen) for quick checks which is great as long as you use <1ug per well

Then we use the bioanalyser to determine integrity

-John Buckels-

hi - dodgy i know, but i just run on a normal agarose (about 2% with etbr) - gives you an idea if it is degraded or not.....should get 2 - 3 discrete bands for 28S, 18S and possibly a 5S if your quality is ok - but i nanodrop for conc etc.....

-aussieuk-

runned in TBE and not TAE.

-fred_33-

QUOTE (fred_33 @ Apr 20 2006, 04:10 PM)
runned in TBE and not TAE.

Why TBE and not TAE?

-sisma-

If you don't want to do denaturing agarose, you can do a denaturing PAGE gel, quite quick and easy - just use whatever rig you use for your SDS-PAGE. Needs to be low %age [like 5%] but will give much sharper resolution of ribosomal subunits. As a guide check out
http://www.protocol-online.org/cgi-bin/pro...ache.cgi?ID=845

-Hooly-

Hi,

i do agree with aussieuk normal agarose gel shud be fine as long as u want to know whether ur RNA is fine (i.e. not degraded). so its fast simple and works well....!

-GeneDoc-

first time heard about E-gel. what that????????? sad.gif

-T. reesei-

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