Autoclaved MOPS: is it supposed to be yellow?
deborah_hailstones_at_emai at smtpgwy.agric.nsw.gov.au
Sun Mar 1 17:26:16 EST 1998
I have never used DEPC treatment in mammalian RNA preps, seeing it as
one of those 'safety nets' that encourage slackness (like antibiotics
in TC....). I have found that if you are suitably meticulous and
cautious it's not necessary to use DEPC, and that way you can also
avoid exposing yourself to (yet another) toxin....
Having said that, my MOPS (in RNA-quality water, alone) ALWAYS goes
yellow after autoclaving and, if made up and then filtered rather than
autoclaved will eventually turn yellow with RT storage.. oxidation,
presumably.... yellow MOPS is definitely fine to use, no worries!
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Subject: Autoclaved MOPS: is it supposed to be yellow?
Author: <bburkho at emory.edu > at smtpgwy
Date: 2/28/98 2:58 AM
I'm trying to set up an RNA gel for a Northern blot.
The protocol I'm using, Current Protocols in Molecular Biology 4.9.1
-4.9.8, gives the recipe for MOPS buffer (10X: 0.4 M MOPS, 0.1 M Sodium
Acetate, 0.01 M EDTA).
I made the solution, added DEPC to treat for remaining RNases, then
autoclaved it. Now my MOPS buffer is yellow, so I'm wondering if
something is wrong.
Is this the correct way to make the buffer, or should I be making it
with water which has previously been treated with DEPC and autoclaved?
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