pyrophosphate generation in PCR - (Sep/17/2008 )
I'm a little confused about how pyrophosphate is generated during the pcr reaction. Is it when the dNTPs bind and start extending the primer? Can someone explain this please?
Is pyrophophate generated even doing nonspecific binding?
pyrophosphate is released which each addition of a dNTP to a growing poly nucleotide chain. So for each nucleoside added to a growing polynucleotide chain you will get one pyrophosphate molecule released.
Hi, thanks for replying...
So theoretically this means then that if I increase the concentration of Mg ions in the reaction then
1) there could be non-specific binding (of polymerase to the remplate) and therefore more non-specific product formation due to the increase in Mg ions
2) Even though the products formed as a result of this, would be primer dimers, it would still generate pyrophosphate wouldn't it due to the few additions of dNTPs?
3) and therefore the increased level of Mg ions could also form Mg-pyrophophate precipitates with the generated pyrophophate isn't it?
Have I got this right?