DNA cleanup - RNAse free

Rich Dudley rdudley+ at pitt.edu
Mon Nov 9 09:56:24 EST 1998


Jens Mollerup wrote:

> Hi Netters,
>
> I need to remove RNA contamination of lambda DNA samples. RNAses are not
> to be used since we will use cRNA (in vitro transkription) later in the
> procedure. Off course we can run an agarose gel and purify from that,
> but there might be an easier way. We have tried to precipitate RNA by
> MgCl2 or LiCl2 - it doesnt work! Does anyone have an idea.
>
> Thanks in advance
> M.Sc. Jens Mollerup

I use RNase to remove any RNA after a prep, followed by phenol:chloroform
extraction, etc.  Our invitro transcription protocol starts with a
linearizing digest and the a proteinase K treatment, whic should remove any
remaining RNase.

Good luck!

rich

--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
http://www.cbp.pitt.edu
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