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RNA gel electrophoresis for integrity check - denaturing or non-denaturing??

RNA gel electrophoresis denaturing gels formaldehyde gel RNA integrity RNA gel

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#1 Neanderthal

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Posted 07 March 2013 - 10:12 PM

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

#2 pcrman

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Posted 07 March 2013 - 11:30 PM

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.

#3 Neanderthal

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Posted 07 March 2013 - 11:37 PM

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

#4 pcrman

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Posted 08 March 2013 - 08:40 AM

You mean loading dye? Yes.

there are a few other posts on this topic:
http://www.protocol-...osts/10527.html
http://www.protocol-...osts/12602.html
http://www.protocol-...osts/10844.html

#5 Neanderthal

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Posted 11 March 2013 - 11:08 PM

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

#6 Trof

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Posted 12 March 2013 - 01:22 AM

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

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#7 Neanderthal

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Posted 12 March 2013 - 09:06 PM

Thanks Trot for the link. It is really helpful.Posted Image





Also tagged with one or more of these keywords: RNA gel electrophoresis, denaturing gels, formaldehyde gel, RNA integrity, RNA gel

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