What are the best conditions to store RNA molecules in? -- Temperature --> I hav

Question

What are the best conditions to store RNA molecules in?

-- Temperature --> I have found that a temperature of -800C is the ideal temperature as stated by some references, and I have also found -200C. I am confused.

-- pH --> I know that it is best to store RNA in non basic conditions so the RNA does not go through hydrolysis (breaking of phosphodiesterbonds).

What are some more conditions to consider?

i.e. salt presence, solvents, amount of RNA, concentration...

Can you please explain why for each condition. Thank you

Explanation / Answer

About temperature i will clear your confusion.If we use DEPC treated water (RNase free) to dissolve the RNA,we can store it in -80C but if we generate cDNA because it is always better to generate the cDNA for the same and store it in -20C in aliquots and main vials in -80C,this is done to avoid freeze thaw cycles.Storing RNA in aliquots is recommended since freezing and thawing repeatedly can degrade our samples.This practice is recommended because cDNA would be more stable for storage that RNA.

DEPC water can inhibit reverse transcriptase PCR reaction,thats why It is always better to dissolve DNA/RNA in Nuclease free water and store them at -80 degrees in small aliquots for long time storage and if you want to generate cDNA you can use one aliquot as discussed earlier.

About pH you rightly mentioned.RNA can be stored in a buffer solution of 1 mM Sodium Citrateand a pH of 6.4.

For Long term storage, precipitate the RNA with ethanol and salt and store the pelleted RNA.Ethanol can be used for storage.

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