Hi there. I'm planning to do an RNA extraction for seeds of the Arabidopsis plant. Currently, I have a saturated phenol solution (pH 6.6). How should I prepare the acid phenol solution from the saturated phenol solution?
To solid phenol add RNase-free water until there is a layer of water on top of the phenol.
Heat new bottle (500g) to 65oC, crack lid. Add 100 ml of RNase-free water.
Mix and let cool. Add about 100 ml of water until a little water remains on top of phenol so that is it completely water saturated. Aliquot into 50 ml tubes and freeze at -20oC.
I remember having it done with Acetic Acid (RNAse free) in a similar way as described by Apoorva .. Because downstream steps in the RNA isolation protocol involve using Ethanol or Isopropanol (better this one, as less volume is necesssary), then alcohol precipitation of your RNA is easy considering that the environment is acid and can be precipitated along with RNA in a sodium acetate-total RNA precipitate. So acetic acid makes sense to me for these reasons. Treated phenol pH has to be 5.0 to be considered saturated with acetic acid. It's painful. I prefer to buy it as phenol is toxic.
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