DNA sterilization before transfection

Chris Boyd chrisb at hgu.mrc.ac.uk
Mon Aug 2 05:08:02 EST 1999


Dr. Robert Gay (r.gay at unsw.edu.au) wrote:
: I'm looking for an alternative way of sterilizing DNA before transfection.

: We currently prepare our DNA with QIAfilter columns and get good yields.
: However before transfection we reprecipitate in 100% EtOH and wash in 70%
: EtOH finally resuspending in sterile water in a TC hood.  However, it
: doesn't matter how careful we are, how long we centrifuge for or at what
: temperature we centrifuge we alway loose most if not all of our DNA !

: Any advice or alternative methods for coming up with transfection quality,
: sterile DNA would be greatly appreciated.

Um, what's wrong with incubating the DNA at 80 degrees or higher for an
hour or so?  (This is a genuine question). Surely this will get rid of
virtually everything alive but spores?  I can't think it would be worse
than EtOH precipitation in this regard anyway.

Best wishes,
-- 
Chris Boyd                      | from (but not \  MRC Human Genetics Unit
Christopher.Boyd at hgu.mrc.ac.uk  | on behalf of) /      Crewe Rd, Edinburgh
http://www.hgu.mrc.ac.uk/Users/Christopher.Boyd          EH4 2XU, SCOTLAND



More information about the Methods mailing list