cloning of PCR products - errors?
roland.hubner at ua.ac.be
Fri Jun 29 05:36:43 EST 2001
> Absolutely. Dropping the [dNTP] to a quarter of recommended values
> helps - it brings it into the Km of the enzyme after all.
I always thought that this was just to keep fidelity higher & would
have expected LOWER product yield with 1/4th concentration, albeit I
observed myself one day same product yield with half amount (was Tfl
PCR, other concentrations were not modified)...
Hmmm, the Km of HiFi-thermopols is different & often more (free?) Mg++
is recommended, but [dNTP] is higher too...
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