71975-M

Sigma-Aldrich

KOD Xtreme Hot Start DNA Polymerase

Ultra high fidelity dnapolymerase designed for the most challenging PCR applications including: crude sample PCR, amplification of long strand and highly GC-rich DNA templates.

NACRES:
NA.85

storage conditions

-20C

Quality Level

manufacturer/tradename

Novagen®

storage condition

OK to freeze

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. Our 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.
The KOD Xtreme<TMSYMBOL></TMSYMBOL> Hot Start DNA Polymerase* kit is an optimized PCR enzyme for the amplification of long or GC-rich DNA templates, including mouse tail-tips, plant tissues and synthetic biology applications. The unique formulation enables you to amplify directly from minimally processed samples; allowing highly efficient crude sample PCR target amplification. The system includes an ultra high fidelity KOD DNA polymerase complexed with two monoclonal antibodies to permit hot start thermocycling, along with specially formulated 2X buffer. KOD Xtreme Hot Start DNA Polymerase quickly and accurately amplifies genomic and phage/plasmid DNA targets up to 24 and 40 kbp, respectively. KOD Xtreme Hot Start DNA Polymerase successfully amplifies challenging DNA templates with up to 90% GC content. Each kit provides 200 U KOD Xtreme Hot Start DNA Polymerase, an optimized buffer and dNTPs sufficient for 200 amplification reactions. The polymerase produces blunt-ended DNA products suitable for cloning with the Novagen Perfectly Blunt<TMSYMBOL></TMSYMBOL> and LIC Vector Kits.
*Manufactured by Toyobo and distributed by EMD Millipore. Not available from Merck KGaA, Darmstadt, Germany 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.

Features and Benefits

  • Optimized for the highest PCR success rate, even with the most difficult targets.
  • 10X higher fidelity than Taq blends
  • Efficiently amplifies up to 90% GC-content DNA templates
  • Amplifies genomic targets up to 24 kbp
  • Amplifies phage/Lambda targets up to 40 kbp
  • Eliminates mispriming and primer-dimer formation
  • Convenient ambient-temperature setup compatible with automation

Components

  • 1 × 200 U KOD Xtreme<TMSYMBOL></TMSYMBOL> Hot Start DNA Polymerase
  • 3 × 1.7 mL 2X Xtreme Buffer
  • 2 × 1 mL dNTPs (2 mM each)

Warning

Toxicity: Irritant (B)

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.

Legal Information

NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany

storage_class_code

10 - Combustible liquids

WGK Germany

WGK 1

Certificate of Analysis

Certificate of Origin

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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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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