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Acid Guanidinium and 2-ME

dkim at unm.edu dkim at unm.edu
Tue Aug 24 09:48:23 EST 1993

Hi again.  I was looking at an ad for USB's Lysate RNAase Protection Assay.
This uses the same lysis procedure as in the AGPC RNA prep, except that there
is no 2-mercaptoethanol added.  I have tried using AGPC lysates for solution
hybridizations and they do not work, maybe the mercaptoethanol is the reason.
Why is there mercaptoethanol at all?  I think, without reducing agents, RNAse
is inhibited in GnSCN down to about 2 M, and with reducing agents, inhibition
is complete down to about 1 M.  Is this the only reason for using 2-ME?  Can I 
get away with omitting it in my regular RNA preps if I keep the GnSCN conc. 
above 2M?  I'd like to try doing Poly(A)+ selection in the crude lysates if
I can get away with it.
Thanks for advice.  Daniel Kim

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