Phenol Protocol

Curt Ashendel ashendel at
Tue Jan 28 12:22:38 EST 1997

On 28 Jan 1997 05:54:08 -0800, Lou Cantolupo  <CANTOLUPO at GENVEC.COM > 

>Does anyone here have a good, reliable protocol for making buffered
>phenol?  I currently use B-M phenol with 8-HQ added, buffered twice with
>untitrated Tris base, then twice with Tric-Cl 8.0.  This works well, but
>takes a good half day to do due to the separations when making 500mL at
>a time.  Thanx in advance....

We use a large sepratory funnel. It speeds up the process with a 
significant increase in the risk of injury to the person doing the 
work (Wear safety glasses and gloves!). It only takes an hour or 
so. Then again, we don't usually make 500 ml at a pop anymore. I don't 
see how you can reduce it to less than three extractions, but you 
probably could get away with just one untitrated tris base 
equilibration as long as the tris base was concentrated enough. 

Curt Ashendel
Purdue University, West Lafayette, IN
ashendel at

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