I would like to know if bacterial RNA in a 1.5 mL 2%SDS solution can be degraded.

Comes from a pellet of pure liquid culture.

  • $\begingroup$ I now that RNA should be freezed in RNAse free water or PBS but this was done accidentally. $\endgroup$
    – biotech
    Mar 27 '15 at 12:45
  • $\begingroup$ I will just store it for 1 week before proceeding to RNA extraction. $\endgroup$
    – biotech
    Mar 27 '15 at 12:49
  • $\begingroup$ It's fine. Your RNA should be stable in the current state. Aside from the fact that it's frozen, meaning that any ribonuclease activity will be minimal (if any), the SDS will have denatured some of the RNase too. However, the detergent (SDS) might impair some of your downstream activities (especially reverse transcription and PCR), so be sure to clean up the RNA well. I would suggest Qiagen columns, and doing the additional wash steps. $\endgroup$ Mar 28 '15 at 11:25

I think it is relatively unlikely that the RNA will degrade under these conditions. For the future, I would handle this differently:

  1. You can centrifuge the cells and snap freeze them in an appropriate buffer in liquid nitrogen and then store them at -80°C. I would not freeze the dry pellet, as these are often hard to re-dissolve after freezing.
  2. Alternatively, you centrifuge them, break them up with Trizol (or whatever you use for this purpose) and freeze them in this solution. This will be safer as it completely denatures all proteins including all nucleases.

I would choose (and have done this already several times with cell samples that I couldn't process without problems) the second method.


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