Denaturing Agarose gel for RNA - (Apr/19/2006 )
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????
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,
but if you run on E-gel (like 2%), then you could also see the integrity of your RNA sample.
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
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.....
runned in TBE and not TAE.
Why TBE and not TAE?
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
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....!
first time heard about E-gel. what that?????????