Lisa J Brunet
brunet at uclink2.berkeley.edu
Thu Aug 25 14:26:43 EST 1994
Janet E. Rubin (jrubi02 at cc.emory.edu) wrote:
: I read two postings regarding the necessity of DTT for RNAsin activity,
: and I seem to have lost them, plus I had never heard of this. Can anyone
: clarify this for me?
: J. Rubin
If you read the Promega literature that comes with RNasin, it will state
that DTT is required for activity. Without DTT, the protein breaks down
and releases RNase. They suggest adding DTT to the RNasin stock every now
and then to replenish what gets lost with every opening of the tube. I
believe the necessary concentration of DTT is 5 mM but I may be mistaken.
By the way, this is why it is important to add DTT to an in vitro
transcription reaction before you add the RNasin.
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