PCR cleanup before digest

newsnet customer customer at newsnet.com
Thu Mar 30 05:15:04 EST 2006


> newsnet customer <customer at newsnet.com> penned the following literary
> masterpiece:
> >Been told to do a PCR cleanup before doing a restriction digest because
> >the salt (MgCl2) in the PCR mix can affect cleavage efficiency. Anyone
> >like to comment?
>
> 1.5mM Mg in a PCR will have negligible affect on an RE digest using 10mM
> Mg.
>
> I would be more concerned with any dNTPs and polymerase remaining active
> i.e. you cut with an RE giving a fillable overhang and the polymerase
> WILL fill it back in using the dNTPs left from the PCR.
>
> Duncan

Well answered.

ST




More information about the Methods mailing list