DNA sterilization before transfection
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.
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
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