Fishing DNA with Dynabeads !!

Csaba Kiss csakis at ki.se
Fri Apr 6 12:40:05 EST 2001


The problem is that it is not one DNA fragment but lots of it. It is
basically a cDNA library that I want to completely digest. Gel purification
is not really an option, since the starting material is a smear and the
resulting digestion is also a smear, although smaller mean size.


"Susanne Rohrer" <srohrer.nospam at immv.unizh.ch> wrote in message
news:3ACD9D23.AE05C9BB at immv.unizh.ch...
> I don't think this will work. think about the melting temperature of a
> tetramer:  between 8 and 16 °C. theoretically. What exactly do you want to
do
> with it?
>
> Csaba Kiss wrote:
>
> > Hi!
> > I want to separate DNA spieces by their overhang
> >  I have to digest my DNA sample. The restriction enzyme gives a 4 bp
> > overhang after digestion. It is absolutely critical that I separate my
> > digested DNA from the undigested. I thought about using dynabeads to
> > separate the digested DNA from the undigested using a 4-bp oligo hanging
off
> > the beads. This oligo can be biotynylated and attached to streptavidin
> > beads. My main worry is if the 4-bp long hydrogen bond between the oligo
and
> > the digested DNA is strong enough to be able to wash away the rest of
the
> > DNA.
> >
> > Csaba
>





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