RNA gels: Autoclaving MOPS buffer?

Tracy Aquilla aquilla at salus.med.uvm.edu
Tue Aug 9 17:29:06 EST 1994


In Article <Aug4.024856.42670 at acs.ucalgary.ca>, ktetro at acs.ucalgary.ca
(Kelly Tetro) wrote:
>In article <1994Aug3.102005.43908 at yogi> herzer at urz.unibas.ch writes:
>>I have read a few protocols on RNA formaldehyde agarose gels.  Maniatis
>>says to sterile-filter the MOPS buffer, another protocol does not say at
>>all (just gives the concentration), and one I got out of FOCUS says to
>>autoclave the 10x MOPS buffer (which turns a nice dark yellow).  Does
>>someone out there have any experience with autoclaving MOPS buffer? 
>>The protocol I'm following looks good, but the yellow MOPS is a bit strange
>>since some others say that the MOPS buffer is bad when it turns yellow.
>>Aughhhhhh!!!!!!
>>               Pete
>>P.S. the paper I'm refering to is called "Northern Blotting: Efficient RNA
>>Staining and Transfer" by Fourney, Miyakoshi, Day, and Paterson ( FOCUS 10:1)
>>I don't have the date here.
>>
>
>Pete, I don't sterilize my MOPS buffer (the one from Maniatis)
>and have gotten success (knock on wood and pray not to offend the
>gods of science). I do make up the buffer in a sterile bottle and
>use sterile reagents as much as possible. Any suggestions on why
>it would need to be sterile? Won't RNAases pass through a filter
>and I thought RNAases weren't inactivated by heat?
>
>Hope this helps:
>
>Kelly Tetro

I usually autoclave things to make them sterile, including MOPS buffer. If
you don't, you risk getting something growing in there, and that is not
always obvious until it's too late. True, RNAses will not be affected (much)
by heat sterilization, but if something starts to grow, it can produce
plenty of RNAse.
Tracy Aquilla, Ph.D.
Dept. of Molecular Physiology and Biophysics
University of Vermont, College of Medicine
aquilla at salus.med.uvm.edu



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