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HotStarTaq PCR Handbook

(with 3 mM MgCl2), and 400 M each dNTP.

Quality Control

Enzyme: (See quality-control label inside kit lid for lot-specific values.) Unit assay: Sonicated herring-sperm DNA (12.5 g) is incubated with 0.010.1 units of HotStarTaq DNA Polymerase in assay buffer (25 mM TAPS [tris-(hydroxymethyl)-methyl-aminopropane- sulfonic acid, sodium salt], pH 9.3 at 20C; 50 mM KCl; 2 mM MgCl2; 1 mM DTT; 200 M of each dNTP; 100 Ci [-32P] dCTP) at 72C for 30 minutes. The amount of incorporated dNTPs is determined by precipitation with trichloroacetic acid. HotStarTaq DNA Polymerase is activated by heating for 3 hours at 80C prior to activity measurement. Amplification efficiency assay: The amplification efficiency is tested in parallel amplification reactions and is indicated under Amp. PCR reproducibility assay: PCR reproducibility and specificity are tested in parallel amplification reactions. The reactions must yield a single
specific product. Exonuclease activity assay: Linearized plasmid DNA is incubated with HotStarTaq DNA Polymerase in PCR Buffer. Exonuclease activity per unit of enzyme is indicat
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