Annealing Temp Primer Calculator How Do I Find A Good Annealing Tp For A PCR Cycle? Why The Primers Don't Melt At The Extention Tp That Is> Tm?

How do I find a good annealing tp for a PCR cycle? Why the primers don't melt at the extention tp that is> Tm? - annealing temp primer calculator

1 I know) that the annealing temperature should be lower due to the Tm of the primers in the Tm of the primer will melt, we have ignited us for a temperature of annealing below the Tm, so that the first model (TM, if choose) to 60 ° C, annealing temp must be 55 to 59. Ok. Thant), but then the temperature at 72 C, by extension, why the caps do not melt at this temperature (which is no longer their TM.
2) in the laboratory, in real time using primers SIBER Green C 45, MC and the cycle of the annealing temperature is 55 ° C are used and have good result, "How?" If I can help someone would understand it greatfull thank you
Sandra