DNA crosslinking on nylon memberane

Tracy Chow via methods%40net.bio.net (by ttchow from gmail.com)
Thu Jul 17 11:04:33 EST 2008


Hi Rory,

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?

THANKS!!!

Tracy

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
> to
> recall several occasions where, using charged membranes, crosslinking was
> omitted but didn't seem to make any discernable difference...
>
> HTH,
>
> - 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
> > _______________________________________________
> > Methods mailing list
> > Methods from net.bio.net
> > http://www.bio.net/biomail/listinfo/methods
>
> _______________________________________________
> Methods mailing list
> Methods from net.bio.net
> http://www.bio.net/biomail/listinfo/methods
>


More information about the Methods mailing list