Southern Blot

peter via methods%40net.bio.net (by peter.ianakiev from gmail.com)
Thu May 24 09:35:36 EST 2007


On May 24, 2:50 am, "Anne" <Anne.Roess... from klinik.uni-regensburg.de>
wrote:
> By PCR? You cut the nylonmembrane and run a PCR with it or how do you do it?
>
> "peter" <peter.ianak... from gmail.com> schrieb im Newsbeitragnews:1179930831.833752.233410 from g4g2000hsf.googlegroups.com...
>
> > On May 23, 8:29 am, "Anne" <Anne.Roess... from klinik.uni-regensburg.de>
> > wrote:
> >> Hello,
> >> does anybody know if it is possible to get the DNA out of the
> >> nylonmembrane
> >> after crosslinking ? I have to know the sequences next to the binding
> >> site
> >> of my sensor.
>
> >> Thanks a lot
> >> Anne
>
> > Yep, by PCR....

That's right Anne - cut the membrane with your fragment on a small
pieces and then use it in a PCR with your favorite primers. Just be
careful to wash out all the SDS which will inhibit PCR reaction even
at very low concentrations. Also for "crosslinking" might be better if
you just use NaOH rather than UV, but that will not interfere with the
PCR if your amplicon is small.
Good Luck!
PS. Sorry to the group for the multiple posts earlier - it is now
obvious that I am using Windows PC....



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