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71086 KOD Hot Start DNA Polymerase

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71086
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          Description
          OverviewPCR involves replication of a DNA template by a thermostable DNA polymerase. The processivity, specificity, and fidelity of the polymerase enzyme used can influence the efficiency, reproducibility, and yield of the PCR reaction. High-fidelity PCR, utilizes a DNA polymerase with a low error rate and results in a high degree of accuracy in the replication of the DNA of interest. Fidelity is critical when accurate sequence amplification of the gene target is needed, for example, when direct sequencing or cloning for downstream protein expression. Unwarranted mutation could severely impact your studies. Our analysis has shown that KOD enzymes are an easy choice for fast, accurate and high-yielding PCR. EMD Millipore's molecular biologists work to develop and formulate polymerases offering the highest specificity, fidelity and yield during PCR amplification. In addition, optimized buffer compositions, convenient master mixes and cycling parameters provide additional ease of use and data reproducibility. 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 amplifies genomic DNA templates up to 21 kb including GC-rich genes for PCR applications. 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.

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          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 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.

          Catalogue Number71086 Brand Family Novagen®
          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
          ReferencesMizuguchi, 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.
          Product Information
          Unit of DefinitionOne 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 [<Sup>3</Sup>H]dTTP), and 150 µg/ml activated calf thymus DNA.
          Components
          DeclarationManufactured by Toyobo and distributed by EMD. 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.
          Quality LevelMQ200
          Applications
          Biological Information
          Physicochemical Information
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          Product Usage Statements
          Storage and Shipping Information
          Ship Code Shipped with Blue Ice or with Dry Ice
          Toxicity Standard Handling
          Storage -20°C
          Do not freeze Ok to freeze
          Packaging Information
          Transport Information
          Supplemental Information
          Specifications
          Global Trade Item Number
          Catalogue Number GTIN
          71086-3 07790788053017
          71086-4 07790788060268

          Documentation

          KOD Hot Start DNA Polymerase Certificates of Analysis

          TitleLot Number
          71086

          References

          Reference overview
          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.

          Brochure

          Title
          High fidelity gene amplification
          PCR Protocols and Guides - Simplify your gene discovery
          The Complete Molecular Biology Toolkit - Expert workflow solutions from DNA cloning to protein expression

          Citations

          Title
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        • Brock F. Binkowski, et al. (2005) Correcting errors in synthetic DNA through consensus shuffling. Nucleic Acids Research 33, e66.
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        • User Protocols

          Title
          TB341 KOD Hot Start DNA Polymerase

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