DNA crosslinking on nylon memberane
(by ttchow from gmail.com)
Thu Jul 17 11:04:33 EST 2008
Thanks for your suggestion...really appreciate.
May I ask if you use Hybond-N+ or Hybond-XL (also positively charged)
membrane? May I ask for the conditions you dry the membrane (room
temperature or vacuum)? How long do you *dry the membranes *prior to
hybridization? How long do you *pre-hybridize* prior to probe incubation?
On Wed, Jul 16, 2008 at 7:34 PM, Rory O'Brien <
rory.obrien from stonebow.otago.ac.nz> wrote:
> Hi Tracy,
> I think DNA binds pretty strongly to positively charged membranes. I seem
> recall several occasions where, using charged membranes, crosslinking was
> omitted but didn't seem to make any discernable difference...
> - Rory
> > -----Original Message-----
> > From: methods-bounces from oat.bio.indiana.edu
> > [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of TC
> > Sent: 17 July 2008 08:40
> > To: methods from magpie.bio.indiana.edu
> > Subject: DNA crosslinking on nylon memberane
> > Hello,
> > I am trying to crosslink DNA to nylon membrane for southern
> > blots. Besides traditional UV crosslinking (may induce DNA
> > damage) and baking (causes DNA to denature), are there any
> > other way to crosslink DNA onto nylon memberanes? For my
> > purpose, I hope minimize DNA damage and keep that DNA in
> > native condition for the Southern blots.
> > THANKS~!
> > Tracy
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