cloning of PCR products - errors?

Wolfgang Schechinger wolfsc at ibms.sinica.edu.tw
Wed Jun 27 10:33:33 EST 2001


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
Academia Sinica
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Tel +886-2-2789-9152
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e mail wolfsc at ibms dot sinica dot edu dot tw
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