Hey, why don't you try a MOP gel? you will get good band separation since your RNA will be completely denatures. It a simple protocol, google it. Hope this helps

RNA + loading buffer + as much formamide as you can squeeze in (aim for 60% or more), heat to 65 for 5mins (65 degrees is about the highest you can heat RNA before you get heat-mediated degradation), crash cool on ice, load on a gel as normal.

This essentially maximally denatures the RNA before loading, so you don't need a denaturing gel.

thanks for your answer but we don't have instruments to check integrity such as Agilent bioanalyzer.

john hildyard, is it necessary to heat sample and add the formamide? because I read this article http://www.ncbi.nlm.nih.gov/pubmed/22222980, also in researchgate discussion while using agarose gel they didn't suggest use of heat or addition of formamide.

i wanna make a fast protocol to check the RNA integrity.

thanks for your help.

Hi, I don't think it is necessary to heat your sample if you are only to check the integrity. We make 1% agarose gel and use Etbr, and the bands of 25S/18S rRNA along with 5S are clearly visible.This is just to check the quality of in vivo RNA.Your procedure sounds fine to me.

Sarah Naiyer

thanks for your help, do you use 1X TAE or 0,5X TAE? 

We use 0.5X TBE in DEPC water but 1X TAE would be fine..

thaks for your help

Adham Sabri,

here is an old conversation I had on that topic.

https://www.researchgate.net/messages.NotificationCenter.html?category=messages&itemId=270887799

Best regards,

thanks Jennifer Taillefond but i cant access your discussion

hello

i tried the following protocol :

1-RNA 1ug+2ul of 6X dye

2-load on 1% gel

3-electrophersis for 30 min at 100v

i got the following results

what do you think about the gel pic?