Error Prone PCR

IndiraShukla Indira_Shukla at brown.edu
Thu May 25 15:27:03 EST 2000


In article <392C05BA.4710D250 at biocomp.unl.edu>, Chris LaRosa
<clarosa at biocomp.unl.edu> wrote:

> Simon Margerison wrote:
> > 
> > Hi
> > 
> > A colleague of mine is interested in performing some error prone pcr.
> > Does anyone know any good methods or reagents for this. Any help would
> > be appreciated, either by direct email to
> > 
> > ruth.featherstone at camb-antibody.co.uk
> > 
> > or on the board.
> > 
> 
> Try using a higher concentration of dNTPs..... this will increase the
> errors.  I know that a lower concentration of dNtps will give lower
> errors.
>  
> I am speculating here: limit the concentration of 3 of the dntps and
> increase the concentration of the remainder.  This should cause more
> errors.  How much so?   Try a range of concentrations....
> 
> I make these comments,, haveing never done such an experiment, so take
> them with a grain of salt.


Try using polymerase which has the proofreading part missing. Such enzymes
are sold commercially by various companies. I have used one such enzyme
with success.






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