PCR 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 XL DNA Polymerase* is an optimized blend of KOD DNA Polymerase and a mutant form of KOD that is deficient in 3′→5′ exonuclease activity (Nishioka 2002). This enzyme mixture is designed for reliable amplification of long, complex targets with robust yield and high accuracy. It can also be used for incorporation of derivatized dNTPs in PCR amplicons (Sawai 2002, Sawai 2001). KOD XL DNA Polymerase generates a mixture of PCR products with blunt and 3′-dA overhangs, suitable for cloning with Novagen Perfectly Blunt®, AccepTor™, and LIC Vector Kits.
Source
Recombinant Thermococcus kodakaraensis KOD1 DNA polymerase expressed in E. coli (wild type and exonuclease-deficient forms)
Concentration
2.5 U/µl
Endonuclease
None detected
Nicking activity
None detected
Amplification effiency
Functional PCR
Storage
–20°C
*Manufactured by Toyobo and distributed by Novagen. Not available from Novagen in Japan. Licensed under US Patent Number 5,436,149 owned by Takara Shuzo Co., Ltd.
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 Number71087 Brand Family Novagen® Features and benefits
Ideal for amplification of large DNA fragments from purified DNA or crude samples
Amplifies DNA templates up to 30 kbp
Successfully amplifies GC-rich sequences
Efficiently incorporates derivatized dNTPs
References
References
Nishioka, M., et al. 2002. J. Biotechnol.88, 141. Sawai, H., et al. 2002. Bioconjugate Chem. 13, 309. Sawai, H., et al. 2001. Chem. Commun. 24, 2604.
Product Information
Unit of Definition
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 [<Sup>3</Sup>H]dTTP), and 150 µg/ml activated calf thymus DNA.
Components
Declaration
Manufactured by Toyobo and distributed by Novagen. Not available from Novagen in Japan. Purchase of this product is accompanied by a limited license to use it in the Polymerase Chain Reaction (PCR) process for research use in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., an authorized thermal cycler.
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