I was confused about phenol while searching it in the context of PCI solution (phenol:chloroform:isoamylalcohol 25:24:1) recipe.
-First, what is the difference between saturated phenol and equilibriated phenol? For example, is phenol saturated with 1 M Tris (pH 8.0) and phenol equilibrated with 1 M Tris (pH 8.0) the same?
-Second, should I use saturated phenol or equilibriated phenol to make a PCI solution?
-Third, when saturating or equilibriating phenol, the suspension separates into an aqueous fraction and a phenolic fraction. We do not use water fraction but the phenolic fraction. Should we discard the aqueous fraction and mix chloroform: IAA (24:1) with phenol or extract the phenolic fraction in a different vessel and use it?