RNA gel electrophoresis for integrity check - denaturing or non-denaturing?? - (Mar/07/2013 )
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,
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:
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.