How important is it to add exactly 1 volume? - phenol:chloroform (Feb/20/2009 )
I have been wondering how important is it to add exactly 1 volume of Phenol:Chloroform:Isoamyl alcohol when doing DNA extraction....for example, you have 800ul of your solution and you would add 1ml of Phenol:Chloroform (instead of 800ul), would it matter?
no, the exact volume added would not matter. It is more important to phenol/chloroform extract until the interface is clear... whereby nearly all the protein in the sample has been removed.
You need to have AT LEAST that much. More is fine, less is NOT
They just tell you to add 1 volume so you don't overfill your eppi tube....