what the best condition of PCR

[kolena]

Hi everybody, I am really need help.
this first time I work with PCR
I have specific primers that have 70% GC rich of forward
and for reveres primer is 64.8%
How I know the best condition of PCR to amplify my target gene.

[luisillo]

You can calculate the melting temperature (Tm) of your primers with this formula:

Tm = (2ºC x no. of A+T) + (4ºC x no. of G+C)

You can take the average Tm of your primers to start standarizing.

Good luck

[uchiharigardo]

You can simulate the PCR reation in this site: http : / / www . idtdna . com / analyzer / applications / oligoanalyzer /
(/\ erase the spaces on the link, because i'm new here and i cant post links)
Analise the primers once by time looking yours features, the concentration of reagents and others parameters.