Phenol Protocol

[Alexander N. Kukushkin]

>   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....
>
>   Lou Cantolupo
>   Dept. Molecular Virology
>   GenVec, Inc.
>   CANTOLUPO at GENVEC.COM
>
During many years we use successfully the following recipe of self-made
buffered phenol to isolate DNA:
1. Mix up 300 ml of distilled melted phenol and 150 ml of TE, pH 8.0.
2. Add 6.75 ml of 5M KOH and 8-hydroxyquinoline to 0.1%.
This mixture is quickly prepared and has pH 8.0-8.2.
Alex