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I work with RNA and I have some RNase issues. For example I am using RNeasy mini kit and at one step I use 100% EtOH. It does not say it is RNase free or not. Does absolute EtOH inhibits or removes RNase or should I be worried about it?

Addition: I am using eppendorfs directly from the bag. So they supposed to be RNase-free, right?

Thanks.

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    $\begingroup$ If nobody else used your EtOH it should be RNAse free, the same is true for tubes. I used tubes fresh from a new bag without problems. But again: Make sure they are handled with gloves all the time and that nobody else uses them. $\endgroup$
    – Chris ♦
    Feb 23, 2014 at 22:02
  • $\begingroup$ I have my own bag for tubes and I am taking them from the bag and putting in a box and I use that box only in "RNase free area". Would that be OK? EtOH is an 1L bottle and everybody taking aliquots from it actually. I have also my own and just me using it. $\endgroup$
    – golgicik
    Feb 23, 2014 at 22:43

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Ethanol is not a problem, we use it without any further precautions and it seems to be RNAse free. You should take the usual precautions you should take with all chemicals when you work with RNA, never use a spatula or spoon to get anything out of the bottle (unless you want to bake your spatulas all the time), always pour them to avoid any contamination.

We autoclave the Eppis once, this might be an unnecessary precaution. I think you can also buy them certified RNAse free, but I'm not sure if they are actually treated any different or simply more expensive.

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  • $\begingroup$ Thanks. What do you think about glass bottles? For instance, I am preparing RNase-free buffers and putting them clean and autoclaved bottles. Do you think it is ok or should I just use plastic containers? $\endgroup$
    – golgicik
    Feb 24, 2014 at 12:21
  • $\begingroup$ I'd probably clean the glass bottles with DEPC first to get rid of any RNAses. Usually baking at 200-250 °C is the way to go for glass, but the bottles typically have some plastic which would not survive this. I also tend to just reuse the same bottles for identical content, then you don't have to make them RNAse-free again. $\endgroup$
    – Mad Scientist - on strike
    Feb 24, 2014 at 12:26

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