Heat-activatable form of KOD DNA Polymerase for increased specificity and convenient PCR setup

KOD Hot Start DNA Polymerase* is a premixed complex of KOD DNA Polymerase and two monoclonal antibodies that inhibit the DNA polymerase and 3′→5′ exonuclease activities at ambient temperatures (Mizuguchi 1999). KOD Hot Start combines the high fidelity, fast extension speed, and outstanding processivity of KOD with the high specificity of an antibody-mediated hot start. Non-specific amplification is reduced because mispriming events that can occur during setup and initial temperature increase are avoided. In addition, primer degradation during setup at room temperature due to exonuclease activity is effectively inhibited. KOD Hot Start DNA Polymerase generates blunt-ended PCR products suitable for cloning with the Novagen Perfectly Blunt® and LIC Vector Kits.

Features and Benefits:

• Highest accuracy, yield, and processivity of commercially available proofreading DNA polymerases
• Amplifies genomic DNA templates up to 12 kbp
• Amplifies plasmid and lambda DNA templates up to 21 kbp
• Successfully amplifies GC-rich sequences
• Eliminates mispriming and primer-dimer formation
• Convenient room-temperature setup compatible with automation
• Optimal KOD Hot Start Buffer for PCR performance over a wide range of targets

References:
Mizuguchi, H., et al. 1999. J. Biochem. (Tokyo) 126, 762. Kitabayashi, M., et al. 2002. Biosci. Biotechnol. Biochem. 66 (10), 2194. Fujii, S., et al. 1999. J. Mol. Biol. 289, 835.

Source	Recombinant Thermococcus kodakaraensis KOD1 DNA polymerase expressed in E. coli
Concentration	1.0 U/µl
Nicking activity	None detected
Amplification effiency	Functional PCR; inhibition of activity at 21°C verified
Storage	–20°C

*Manufactured by Toyobo and distributed by EMD. Not available from Merck in Japan.

Note: Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser's own internal research. No other patents rights (such as 5' Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California, 94404, USA.

Unit Activity: One unit is defined as the amount of enzyme that will catalyze the incorporation of 10 nmol of dNTP into acid-insoluble form in 30 min at 75°C, in a reaction containing 20 mM Tris-HCl (pH 7.5 at 25°C), 8 mM MgCl₂, 7.5 mM DTT, 50 µg/ml BSA, 150 µM each of dATP, dCTP, dGTP, dTTP (a mix of unlabeled and [³H]dTTP), and 150 µg/ml activated calf thymus DNA.

Components:

•	200 U or 5 × 200 U	KOD Hot Start DNA Polymerase (1.0 U/µl)
•	1.2 ml or 5 × 1.2 ml	10X PCR Buffer for KOD Hot Start DNA Polymerase
•	1 ml or 5 × 1 ml	25 mM MgSO4
•	1 ml or 5 × 1 ml	dNTP Mix (2 mM each)