Buffering Phenol(sounds like 'Kit Wars')

dallas at mcvax4.d48.lilly.com dallas at mcvax4.d48.lilly.com
Thu Jun 16 13:08:05 EST 1994


In article <2t7bmf$5r7 at bigboote.WPI.EDU>, jbagshaw at wpi.WPI.EDU (Joseph C. Bagshaw) writes:
> 1.  Consider the cost of the Tris you will use up and the amount
> of phenol you will lose (some goes into every aq. phase).
> 
> 2.  Consider the amount of time you will spend on this task.
> Your graduate student doesn't count; she has more useful things
> to do.

Heavens, let's not have a graduate student learn something by doing!!

> 
> 3.  Look up the price of buffered phenol in any of a half dozen
> catalogs and compare it with the price of unbuffered phenol.
> 
> 4.  Buy buffered phenol.

Shouldn't throw stones, but when I was in graduate school, we couldn't buy such
niceties.  In fact, I had to distill my (f)enol [for the 'phans'].
> 
> 
Please don't take comments personally.  I just came from a newsgroup with a lot
of 'flame wars'.

Bring on 'kit wars'.

Jim Miller
Lilly Research Labs
Indianapolis, IN

> 
> In article <199406071653.JAA27400 at net.bio.net> MUNGO at UVVM.UVIC.CA writes:
>> We have on occaison found that it can be very difficult to buffer phenol to
>>pH 8.  For reasons nobody here can explain certain lots of high quality phenol
>>are impossible to get to pH 8 even after overnight extraction with Tris base
>>0.5 M unbuffered.  The phenol remains at pH 6.8.  However someone in the lab
>>added a few drops of 10 M NaOH and the two phases then mixed and wouldn't
>>seperate.  Upon addition of 50 mM Tris pH 8 the pahses seperated and the phenol
>>was at pH 8 (sort of...).  Repeated extraction with 50 mM then 10 mM Tris pH 8
>>does not change the pH of the phenol.  While I don't wish to address this
>>rather novel approach to buffering phenol can anyone tell me what is going on?
>>Why does adding NaOH cause the phases to mix ?  Is it the salt ?  Why do we
>>have trouble buffering the phenol in the first place if it easily buffers once
>>the phases mix ?  If the salt is causing the phases to mix why does the phenol
>>remain buffered after repeated extractions with low salt buffer ?  And
>>is this "solution" usable ?
>>Thanks
>>Mungo
> 
> 
> -- 
> 
> **********************************************************************
> Joe Bagshaw                        HAVE GENES, WILL TRAVEL
> Department of Biology and



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