RNA gel electrophoresis for integrity check - denaturing or non-denaturing?? - (Mar/08/2013 )

Dear all,

I would like to know which is the preferred method for checking the integrity of RNA (stored for >1 year in -80) using gel electrophoresis. I want to use this RNA for real-time PCR. Should I do denaturing gels or non-denaturing gels?

In non-denaturing gels. should I use 1x TAE or 1x TBE?

Thanks a lot,

Kart

You can use agarose gel in ATE buffer as used for DNA to check RNA quality. Just make sure your gel system is RNase free.

If I can use TAE buffer that I use for DNA, can I use the same loading buffer also?

You mean loading dye? Yes.

there are a few other posts on this topic:
http://www.protocol-online.org/biology-forums-2/posts/10527.html
http://www.protocol-online.org/biology-forums-2/posts/12602.html
http://www.protocol-online.org/biology-forums-2/posts/10844.html

oh..yes, I meant loading dye. Thanks a lot for the links.

Also, we check the RNA in denaturing conditions using non-denaturing gels, just adding formamide to the loading buffer (so no special preparations).

Thanks Trot for the link. It is really helpful.