cloning of PCR products - errors?

[AvV]

Higher Mg gives more errors in my experience. Lower to 1.5 mM if possible?

Arnoud

""Wolfgang Schechinger"" <wolfsc at ibms.sinica.edu.tw> wrote in message
news:3B3A6E04.24965.25F70CE at localhost...
> The main issue probably is the number of copying cycles
> which depends on the amount of template originally provided. If
> you start with one copy and ampflify it to 1 ug, there should be
> plenty of typos in there...
>
> Any quantitative experience from anyone?
>
> Wolfgang
>
> >
> >
> > Roland Hubner wrote:
> >
> > > In article <3B39CD96.B8AB041 at bbm1.ucm.es>, Sergio
> > > <sergioal at bbm1.ucm.es> wrote:
> > >
> > > > Only 265x10-6 ??... damn... i must be doing something wrong.
> > > > We use to get around 1x10-3 mutations when using Taq...
> > > > almost five times the supposed error rate.
> > > >
> > > > Sergio
> > >
> > >  cycle numbers?
> >
> > 25 cycles
> > MgCl2 2.5 mM
> > dNTPs 0.25 mM each
> > primers  0.5 uM each
> >
> > 95 (1') --- Tm-2 (2') -- 72 (1'/kb)
> >
> > All reagents from Perkin Elmer
> >
> > Sergio
> >
> >
> >
>
>
> [tiss mezzage wahs broduceRd using TYPO GENERATOR zoffwer]
> -----
> Dr. Wolfgang Schechinger
> Lab N233 (c/o Dr. Steve Roffler)
> Institute of Biomedical Sciences
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