71086

Sigma-Aldrich

KOD Hot Start DNA Polymerase

High fidelity DNA polymerase designed for accurate PCR amplification of long strand and GC- rich DNA templates for cloning and cDNA amplification applications.

storage conditions

-20C

Quality Level

manufacturer/tradename

Novagen®

storage condition

OK to freeze

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description



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 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.
Source: Recombinant Thermococcus kodakaraensis KOD1 DNA polymerase expressed in E. coli
Concentration: 1.0 U/l
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.

Packaging

200 u in Fibre case
1000 u in Glass bottle

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 UKOD Hot Start DNA Polymerase (1.0 U/µl)

•1.2 ml or 5 × 1.2 ml10X PCR Buffer for KOD Hot Start DNA Polymerase

•1 ml or 5 × 1 ml25 mM MgSO₄

•1 ml or 5 × 1 mldNTP Mix (2 mM each)

Warning

Toxicity: Standard Handling (A)

Unit 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 [3H]dTTP), and 150 µg/ml activated calf thymus DNA.

Other Notes

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.

Legal Information

Manufactured 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.
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany
Perfectly Blunt is a registered trademark of Merck KGaA, Darmstadt, Germany

storage_class_code

10-13 - German Storage Class 10 to 13

Certificate of Analysis

Certificate of Origin

Jun Wei et al.
Nature, 576(7787), 471-476 (2019-12-13)
Adoptive cell therapy represents a new paradigm in cancer immunotherapy, but it can be limited by the poor persistence and function of transferred T cells1. Here we use an in vivo pooled CRISPR-Cas9 mutagenesis screening approach to demonstrate that, by...
Articles
The purpose of Hot Start PCR is to inhibit the PCR reaction in order to reduce nonspecific amplification, prevent the formation of primer dimers, and increase product yields.
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Protocols
Creating Transgenic Mice using CRISPR-Cas9 Genome Editing
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