RNA affinity column

Ian A. York iayork at panix.com
Thu Sep 16 18:15:00 EST 1999


This is something I'm thinking about as a concept rather than an
experiment for tomorrow, if you see the difference.  Let's say I have an
RNA that I suspect of interacting with some unknown proteins.  One way of
purifying the proteins would be to make an affinity column:  bind the RNA
to the column, pour cell lysates over it, wash, and elute.

Can this be done?  I'm no RNA dude.  The RNA in question is around 500
bases long.  It seems to me that there could be a number of obstacles:
(1) Production of enough RNA.  How much, and how clean, could I get in an
in vitro system?  Or is there a way of production in bacteria that will
allow  ...
(2) Tagging of the RNA; either for identification, or for easy ...
(3) Attachment to the column.  Can the RNA be modified to allow one end to
react, but not the other?
(4)  Elution seems straightforward enough, in concept, if I can get the
other things to work; just denature proteins and wash.

I'm not finding anything relevant in medline, but that might just be
because I don't know the right key words.  I have a vague idea that this
appraoch has been taken for HIV or something.  Any pointers appreciated.

Ian 

-- 
    Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
    "-but as he was a York, I am rather inclined to suppose him a
     very respectable Man." -Jane Austen, The History of England



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