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