Using Phenol - (Jul/31/2007 )
I had a doubt about using phenol, as I have never used it before for DNA extraction purposes.
The Molecular Cloning Lab Manual suggests equilibrating commercially available liquefied phenol and recommends not to use the crystalline form.
Is this procedure necessary?
thanks a bunch
yes, you must equilibrate your phenol. In basic phenol (~pH 8), the DNA is in the aqeous phase. In acidified phenol (~pH 4.5), RNA is in the aqeous phase. Change the pH and you can selective selectively extract either RNA or DNA. Get the pH wrong and you'll extract the wrong thing.
i recently bought a phenol bottle, and it came at pH roughly 6. there was a small bottle with the phenol one, and user was supposed to put it in the phenol to euilibrate at pH 8.
Could you tell me what company did you buy the phenol from. The one that gives you another solution to equilibrate the phenol.
I did not know this that differences in the pH can lead to extraction of different nucleic acids. Good to know it!
it was invitrogen.
I'm not at lab so for precise ref it will be tough but may search you that.