RNA gels: Autoclaving MOPS buffer?

Kelly Tetro ktetro at acs.ucalgary.ca
Wed Aug 3 21:48:56 EST 1994


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



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