cloning the wrong way?

Sergio sergioal at solea.quim.ucm.es
Thu Feb 17 04:26:58 EST 2000



"Richard P. Grant" wrote:

> In article <38AAD3B4.D212C32C at uni-konstanz.de>,
> Frank.Fackelmayer at uni-konstanz.de wrote:
>
> > ALWAYS!!! Especially when using Taq polymerase which is notorious for
> > making mistakes.
>
> Pish.
>
> To the second part of this, anyway.  Drop the dNTP conc to 50 uM (from
> 200 uM) and watch your error rate fall away.
>
> R

Sounds interesting, since i have lots of PCR clonings to do (and i always get a
mutation rate around 2/1000). Do i need to change any parameter in the PCR
conditions if i use 50 uM dNTP?

thanks in advance

Sergio






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