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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?

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  • $\begingroup$ Honestly? Don't make this solution yourself, buy it ready to use from a lab supplier. Phenol is nasty stuff and you produce a lot of waste making the solution yourself. It's not so expensive one could justify handling it more than necessary. $\endgroup$ – Chris Jan 30 at 8:14

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