The innuTaq HOT-A DNA Polymerase provides improved specificity and sensitivity when amplifying low-copy-number targets in complex backgrounds or when prolonged room temperature set up is required. The polymerase activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of nonspecifically annealed primers and primer-dimer formation at low temperatures during PCR setup. The enzyme catalyzes the polymerization of nucleotides into duplex DNA in 5‘ → 3‘ direction in the presence of magnesium. It also possesses a 5‘ → 3‘ polymerization-dependent exonuclease replacement activity but lacks a 3‘ → 5‘ exonuclease activity. innuTaq HOT-A DNA Polymerase requires no prolonged heating or denaturing step. The polymerase inhibiting ligand is quickly released at the increased temperature of thermal cycling. The innuTaq HOT-A DNA Polymerase provides improved specificity and sensitivity when amplifying low-copy-number targets in complex backgrounds or when prolonged room temperature set up is required. The polymerase activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of nonspecifically annealed primers and primer-dimer formation at low temperatures during PCR setup. The enzyme catalyzes the polymerization of nucleotides into duplex DNA in 5‘ → 3‘ direction in the presence of magnesium. It also possesses a 5‘ → 3‘ polymerization-dependent exonuclease replacement activity but lacks a 3‘ → 5‘ exonuclease activity. innuTaq HOT-A DNA Polymerase requires no prolonged heating or denaturing step. The polymerase inhibiting ligand is quickly released at the increased temperature of thermal cycling.