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.
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.
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
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)
*Manufactured by Toyobo and distributed by EMD Biosciences, Inc. Not available from EMD Biosciences, Inc. in Japan