How to make RNase-free glycogen solution?

Chris Seidel seidel at mendel.berkeley.edu
Thu Feb 16 16:55:12 EST 1995


In article <01HMUVW1ZEGI8WWGOC at CCVAX.MMC.EDU>, CHAUDH45 at CCVAX.MMC.EDU
wrote:

> 	I will highly appreciate a method for making a solution of glycogen RNase free. Can DEPC be used ? How to remove residual DEPC?

Usually I phenol extract glycogen when preparing RNase free aliquots
of it, and that seems to work fine. DEPC modifies more than just
polymerases and RNases, it is a strong acylating agent that also
modifies RNA and DNA. I think for most applications this is not much
of a concern. Typically DEPC is removed by autoclaving, which I wouldn't
try with glycogen. However DEPC is so reactive that even at 
room temperature, at the limit of it's solubility in water (40 mM),
it has a very short half life on the order of minutes. So if you'd
like to prepare your glycogen that way (I don't know of DEPC/Glycogen
reactions) you could presumably just let it sit at room temp
for some time period. Someone in the lab here uses it for footprinting
and has to keep ordering it fresh because it goes off so quickly.

Chris



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