Pure recombinant high fidelity DNA polymerase from Thermococcus kodakaraensis KOD1

KOD DNA Polymerase* (formerly KOD HiFi DNA Polymerase) is a recombinant form of Thermococcus kodakaraensis KOD1 DNA polymerase (Nishioka 2001). KOD is a high fidelity thermostable polymerase that amplifies target DNA up to 6 kbp with superior accuracy and yield (Takagi 1997). The enzyme’s 3′→5′ exonuclease-dependent proofreading activity results in a lower PCR mutation frequency than any other commercially available DNA polymerase. The elongation rate and processivity are 5 times and 10 to 15 times higher, respectively, than for Pfu DNA polymerase, resulting in highly accurate and robust yield, in a short reaction time. The enzyme generates blunt-ended PCR products suitable for cloning with the Novagen Perfectly Blunt® and LIC Vector Kits.

Features and Benefits:

• Higher fidelity than Pfu DNA polymerase—excellent for cloning
• Greater yield—extension speed is 2X faster than Taq DNA polymerase and 5X faster than Pfu DNA polymerase
• Higher processivity—sequential nucleotide polymerization is 10- to 15-fold greater than Pfu and Tli DNA polymerases
• Amplifies plasmid and lambda DNA templates up to 6 kbp
• Amplifies genomic DNA templates up to 2 kbp
• No truncated amplification products

References:
Nishioka, M., et al. 2001. J. Biotechnol. 88, 141. Takagi, M., et al. 1997. Appl. Environ. Microbiol. 63, 4504.

Source	Recombinant Thermococcus kodakaraensis KOD1 DNA polymerase expressed in E. coli
Concentration	2.5 U/µl
Purity	> 90% homogeneous by SDS-PAGE
5′ Exonuclease	Less than 2% per unit of enzyme when incubated 1 h at 74°C in a reaction with 5′-labeled l/Sca I digest
Nicking activity	None detected
Amplification effiency	Functional PCR
Storage	–20°C

*Manufactured by Toyobo and distributed by EMD Biosciences, Inc. Not available from EMD Biosciences, Inc. 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:

•	250 U	KOD DNA Polymerase (2.5 U/µl)
•	1 ml	10X Buffer #1 for KOD DNA Polymerase (pH 8.0)
•	1 ml	10X Buffer #2 for KOD DNA Polymerase (pH 8.8)
•	1 ml	25 mM MgCl2
•	1 ml	dNTP Mix (2 mM each)