| | Re: Asking about Hot Start PCR
"Hot-start" is basically a method to prevent non-specific amplification during the early stage of PCR, ie. as the temperature starts climbing up to 94-95 for the initial denaturation.
"Hot-start" DNA polymerases are enzymes that only become active after the initial denaturation. This usually involve the binding of antibodies/beads/etc to the enzyme that inhibits it, and will only dissociate from the enzyme at a hight temperature of 94-95. In other words, no amplification can occur as the PCR machine ramps up the temperature to 95 for the initial denaturation.
You can also run a "hot-start" PCR manually, eg:
1. Leave out a critical component of the PCR, eg Mg or Taq until the PCR machine reaches 94. Then only you add the final component.
2. Set a longer period for the initial denaturation. After the PCR machine ramps up to 94, then only you place your samples on the machine.