How do YOU precipitate your RNA? - (Jan/20/2006 )
I'm in a new lab and as usual: new lab new prayers so to say. Or in other words: every head of lab or scientists is convinced that his and only his method works for real - as many people as many methods it seems.
Now what I started to realize and made me pretty curious too is that even for something as basic as precipitation of RNA there are countless ways to go. Which one do you use and why? Shouldn't there be some perfect method?
What I tend to vote for from a completelly mental excerise but no idea if this gives really a difference is option three (Ethanol and NH4-Acetat pH5.5) as I read somewhere that using Na-Acetat is increasing the salt inside while NH4 is not and having acidid pH helps to decrease the solubility of RNA.
Another thing I got interested in and we could maybe combine: what about cooling before the centrifugation?
Are you just pipetting it on ice together and centrifuge it? Or do you go for the 30' -80°C option? Or 2 hours 4°C? Or overnight 4°C? Or 10' dry ice (which worked marvelous for a 5' race kit, had not believed so short cooling could do the trick)?
Curious about your experiences here, so shoot straight please.
Always keep your RNA on ice as much as possible, or you will end up with none of it (RNase digestion).
Cooling: I tend to go for the dry ice option (the quicker you work with the RNA the less chance of digestion).
One suggestion I would like to make, is that you centrifuge longer than with DNA and centrifuge again after the 70%EtOH wash, as RNA pellets are not as sticky.
Are you just pipetting it on ice together and centrifuge it? Or do you go for the 30' -80°C option? Or 2 hours 4°C? Or overnight 4°C? Or 10' dry ice (which worked marvelous for a 5' race kit, had not believed so short cooling could do the trick)?
Curious about your experiences here, so shoot straight please.

8M LiCl works well