Long Term RNA Storage

[bgrimes]

What is the best buffer to store RNA extracts in if you intend to store it for many years? I know most protocols call for simply eluting in RNase free water and storing at -80, but I can't find much info on very long-term storage.

Ambion recommends TE or a sodium citrate buffer. Does anyone have experience either way using these buffers?

Thanks for any help

[songsong]

I usually store them in 75% ethenal,two years or maybe longer. :-)

[̑̑̑]

i normally store it in a box... (lol jk!), nah, depc treated water at -80 and it seems to be alright at least for few years (the whole of my phd, so 3 years in my case)

[moleculardude]

Originally Posted by ̑̑̑

i normally store it in a box... (lol jk!),

Has anyone dried RNA and stored dry?

I bet this would last very long. What about RNAse inhibitors added?

cheers

[kanzure]

How would buffers be used to cryogenically store RNA?

- Bryan

[danfive]

TE, SSC, and Ultrapure H2O, are all equivalent, in Rnase free tubes (ambion). I would record RNA concentration in a logbook, just in case. May be helpful if you need to troubleshoot after years of storage. Most people I know would not be surprised if low concentration of RNA fails in RT-PCR after a few months in any freezer. Folklore/Lab theory goes that some RNA will be lost to simple sticking to the tube surface.

[danfive]

Originally Posted by kanzure

How would buffers be used to cryogenically store RNA?

- Bryan

Consider a Trizol RNA extraction and final step, the RNA pellet is resuspended in TE or SSC buffer. You can then make a 1:20 (or whatever) dilution and take an absorbance to measure concentration. Freeze your stock RNA.

[danfive]

Originally Posted by bgrimes

Ambion recommends TE or a sodium citrate buffer.

This is so true, Ambion is a big fanatic of RNA in SSC. They even sell you special SSC for RNA storage. I found that it is useless if you are storing very low RNA solutions.