Sv: Sonication of DNA for cloning
ali at image.dk
Wed Feb 28 04:01:58 EST 2001
It is diffucult for me to find suitable restriction endonucleases as the
gene is more than 80 percent AT
Sergio <sergioal at bbm1.ucm.es> skrev i en
nyhedsmeddelelse:3A9BD8D5.3EF379F2 at bbm1.ucm.es...
> I cannot answer your question, but, why don't you clone the whole PCR
> and afterwards subclone it in smaller fragments using restriction
> salanti wrote:
> > I am going to make a small library from a 7kb PCR fragment. I plan to
> > fragment the big PCR product by sonication, then treat it with Klenow.
> > But what happens to the "nicks" in the DNA that will occur from the
> > sonication?
> > In a protocol using DNAse 1 to digest the DNA, they point out that it is
> > important to use a Mn+ buffer instead of Mg+ in order to avoid nicking
> > DNA!! My question is : Is it OK to sonicate DNA for cloning purposes
> > Thank you
> > Salanti
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