DNA crosslinking on nylon memberane

TC via methods%40net.bio.net (by tracyscience from gmail.com)
Thu Jul 17 13:39:49 EST 2008


Hi all,

Thanks for sharing your thoughts.  I hope to study the overhangs of the DNA;
therefore, need to keep DNA in native conditions.

Tracy

On Thu, Jul 17, 2008 at 1:25 PM, Tracy Chow <ttchow from gmail.com> wrote:

>
>
> ---------- Forwarded message ----------
> From: Jose de las Heras <josenet from tiscali.co.uk>
> Date: Thu, Jul 17, 2008 at 4:14 AM
> Subject: Re: DNA crosslinking on nylon memberane
> To: methods from magpie.bio.indiana.edu
>
>
>
> "TC" <tracyscience from gmail.com> wrote in message
> news:mailman.777.1216254116.3533.methods from net.bio.net...
> > 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
>
> Several positively charged membranes claimed to be ok for use without
> conventional crosslinking. Personally I always UV-crosslink out of habit.
>
> I doubt the damage on immobilised DNA is going to give you any trouble, and
> I doubt immobilised DNA on a dry membrane is going to denature with heat...
> and besides, for successful hybridisation you want the DNA to be denatured.
>
> You may try a positively charged membrane to bypass heating/UV.
>
> Out of curiosity, what are you trying to do?
>
> Jose
>
>
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