Autoclaving MES

Michael L. Sullivan mlsulliv at facstaff.wisc.edu
Wed Jan 23 17:20:28 EST 2002


>On Wed, 23 Jan 2002, Michael Witty wrote:
>
>> On 23 Jan 2002, Michael L. Sullivan wrote:
>>
>> > >
>> > >. . . anyone tried this?  Or does it break down.  Mike.
>> >
>> >
>> > I've used MES as a buffering agent in plant tissue culture media before,
>> > using an autoclaved stock and then re-autoclaving the media.  From the pH
>> > adustment prior to the second autoclaving, the MES had certainly retained
>> > buffering capacity, although I couldn't tell you that there was no
>> > breakdown at all.  It is certainly a fairly standard practice to make
>>plant
>> > tissue culture media and autoclave it with the MES in it, but I suppose
>> > that doesn't mean there are no effects on the MES, just that that is how a
>> > lot of people do it!
>> >
>> > Hope this was somewhat helpful.
>> >
>> > Mike
>>
>> . . . Thanks Mike.  I also phoned up the manufacturer: the real guy in the
>> Factory up north who makes it.  He is a chemist and says it should be
>> "stable way above 120 degrees".  I will bravely risk my 20x stock.  But
>> tomorrow.  Regards, Mike W.
>
>. . . but wait!  A quick Google search has turned up
>http://www.fgcz.unizh.ch/Affymetrix_manual.pdf and
>http://www.cancergenetics.med.ohio-state.edu/microarray/protocols/GeneChipAssay
>1.pdf
>which say MES should not be autoclaved.  And furthermore to throw it away
>if it turns yellow (though this is not true because my bright yellow 20x
>MES stock for PAGE is still working fine.  Life is so complex!  Mike.

Are these referring to proceedures similar to what you will be doing (gene
chip stuff?)?  I guess I'd follow their advice if that's the case, although
maybe it doesn't REALLY matter.  An awful lot of proceedures call for doing
things a particular way for no particular reason!

This sort of reminds me of discussions that have taken place here regarding
whether it's OK to use yellow MOPS buffer to run RNA gels.  Despite the nay
sayers, I've used yellow mops for several hundred northern blots with great
results!

The other Mike

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