I have my RNA samples eluted in nuclease-free water. But now I want to store it for a longer period. Can I add isopropanol directly to my samples to store it without doing a second round of TriZol treatment? Will I be able to recover the sample back in this way?
You can store it in rnase free water at -70c or in ethanol or isopropanol directly. Keep it as single time use multiple aliquotes to avoid freeze thawing causing damage to rna. U can also store in trizol or buffer like tris 10mM PH7.
Hello Chandan Kumar Pradhan
For long-term storage, RNA preps may be stored at -70 to -80ºC either in RNase-free water or precipitated in isopropanol. You already have your RNA samples eluted in nuclease-free water. So, you may store it at - 70 to -80ºC for long -term storage as RNA is relatively stable at this temperature. However, to avoid repeated freeze-thaw cycles, it is recommended that frozen RNA samples be stored as multiple, single-use aliquots.
Alternatively, you may store RNA at -70 to -80°C, as aliquots precipitated in isopropanol. Later when there is a requirement, centrifuge the RNA and resuspend in an appropriate RNase-free buffer before use.
Best.
Chandan Kumar Pradhan
For the long term, RNA can be stored in either nuclease-free water or precipitated in ethanol or isopropanol at -70 C as a single-use aliquot to prevent multiple freeze-thaw. Later, when needed, centrifuge the aliquot and resuspend it in the nuclease-free water.
Reprecipitating RNA purely for long term storage is not a great idea. Precipitation isn't 100% efficient, so you'll lose RNA, and then when you redissolve it in future you'll have to spec it again to determine concentration, so you'll lose even more.
More to the point, RNA isn't much less stable in water than it is precipitated, especially at -80. And at -80, RNA in water is stable for _years_.
Store it at -80 as it is. You know how much you have (ng.ul-1), so when you need it in future, you can just get it out and use it.
One thing to note is that freeze/thaw cycles are not great for RNA integrity, so if you're planning to dip in and out of your stored RNA multiple times, it might be better to store it in usefully-sized subaliquots.
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