HotStar PCR SuperMix provides qualified reagents for the amplification of nucleic acid templates by polymerase chain reaction (PCR). The mixture contains anti-Taq DNA polymerase antibody, Mg⁺⁺, dNTPs, and recombinant Taq DNA polymerase at concentrations sufficient to allow amplification during PCR. HotStar PCR SuperMix is supplied at 2X concentration to allow approximately 50% of the final reaction volume to be used for the addition of primer and template solutions. Reagents sufficient for 100 amplification reactions of 50 μl each are provided. Anti-Taq DNA polymerase antibody inhibits polymerase activity providing an automatic hot start and permits ambient temperature setup. Antibody-mediated hot starts improve PCR specificity and yield. Due to specific binding of the antibody, HotStar PCR SuperMix is present in an inactive form and is reactivated after a denaturation step in PCR cycling at 94°C. HotStar PCR SuperMix may be stored at either -20°C or 4°C. Storage at 4°C avoids the necessity of thawing the mix before assembling the PCR. Repeated freeze-thaw cycles might reduce performance or activity.