RNA gel electrophoresis

[sumaBiology]

Hello all,

Does any one could send me the easiest and convenient protocol of RNA gel electrophoresis to check if my isolated RNA is Degraded or not?.

Thanks in Advance.

[r.rosati]

Hello!
The easiest and cheapest way is to run a native agarose gel.
Check the small protocol at the bottom of >this page<, entitled "Native Agarose Gel Electrophoresis of RNA".

Remember:
-All gel plasticware (casting tray etc.) must be thoroughly washed with soap and rinsed in ddH2O (best if mQ-grade water)
-TBE can be autoclaved. In my lab we use 1xTAE prepared from a 50x stock, it gives good results too, but TAE shouldn't be autoclaved. Prepare from mQ-grade water and be clean - remember that autoclaving can reduce the RNAse burden, but can't make contaminated water RNAse-free (and you shouldn't add concentrated DEPC to a solution with amines like Tris).

Roberto

[pidou213]

I think you can make a native gel check ,and if you find the 28S and 18S RNA are intergrity. That tells you your RNA is good.

[sumaBiology]

hi all,
Thanks,I tried with the native gel......which worked fine with me

Thanks again

[.anka]

I isolated RNA from FFPE tissue sample with FFPE RNeasy kit and then checked RNA samples 1% native agarose gel, but there was just one sharp band. What do you think about this?

I appreciated for your helps, thanks...

[brainsnomnom]

21 June 2012 J3T 24h RNA corrected.jpg

Just wanted to post a picture of what my samples look like when I run them on 2% agarose gel with Gel Red dye in the gel, using ~1ng of RNA with 10ul of Ambion RNAqueous loading dye with formalin. The ladder is a 1kb DNA ladder. My bands are wiggly because I am a crappy gel maker and get impatient and take the combs out before they're ready

[brainsnomnom]

Whoa, sorry, meant 1ug.