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71842 Sigma-Aldrich

KOD Hot Start Master Mix

Ready-to-use 2X mixture, containing KOD Hot Start DNA Polymerase, two monoclonal antibodies, ultrapure deoxynucleotides, and reaction buffer with MgSO4, optimized for convenient high fidelity PCR., ready-to-use solution, suitable for PCR

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Properties

Related Categories Hot Start PCR, KOD PCR Products, Molecular Biology, PCR/Amplification
Quality Level   100
form   ready-to-use solution
mfr. no.   Novagen®
storage condition   OK to freeze
storage conditions   -20C
application(s)   PCR: suitable (2X PCR Master Mix for high fidelity)
shipped in   dry ice
  wet ice

Description

General description

KOD Hot Start Master Mix is a ready-to-use 2X mixture optimized for convenient high-fidelity PCR, which 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. It contains KOD Hot Start DNA Polymerase, two monoclonal antibodies, ultrapure dNTPs, and reaction buffer with MgSO4. The mix is ideal for use in high-throughput PCR applications.

Application

KOD Hot Start Master Mix has been used:
• To yield PCR product for T7 endonuclease I (T7EI) assay.
• For PCR amplification of the DNA isolated from Chrysosporium multifidum, a fungus with antimicrobial activity.
• In the PCR amplification of cDNA.
• In two-step PCR for the amplification of the target sequence followed by its insertion into a vector.

Packaging

100 reactions in Plastic ampoule

500 reactions 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 template up to 21 kbp
• Eliminates mispriming and primer-dimer formation
• Convenient ambient-temperature setup compatible with automation
• KOD HotStart® Buffer ensures optimal PCR performance over a wide range of targets.
• Offers high specificity, fidelity and yield during PCR amplification
• Optimized for ease of use and data reproducibility
• Simplifies PCR set-up and saves time
• High consistency with minimal risk of contamination

*Manufactured by Toyobo and distributed by Merck KGaA, Darmstadt, Germany and/or its affiliates (not 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.

Components

•2 × 1.25 ml or 10 × 1.25 mlKOD Hot Start Master Mix

>References:

>Kitabayashi, M., et al. 2002. Biosci. Biotechnol. Biochem.66 (10), 2194. Fujii, S., et al. 1999. J. Mol. Biol.289, 835.

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

Simply add KOD HotStart® Master Mix to an equal volume of sample containing DNA template and primers. The final diluted reaction contains 1U KOD HotStart® DNA Polymerase per 50 μl reaction. The smaller available size provides sufficient master mix for 100 (50 μl scale), or 250 (20 μl scale) reactions, while the larger size is adequate for 500 (50 μl scale) or 1250 (20 μl scale) reactions.

Legal Information

* Manufactured by Toyobo and distributed by Merck KGaA, Darmstadt, Germany and/or its affiliates (not 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.

HOTSTART is a registered trademark of Molecular BioProducts, Inc.

Safety & Documentation

Safety Information

WGK Germany 
WGK 2
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable
Protocols & Articles

Articles

Hot Start PCR

Hot Start PCR is a technique that inhibits Hot Start Taq polymerase activity or the incorporation of modified dNTPs during reaction set up until a heat activation step occurs. Hot Start PCR allows fo...
Keywords: Amplification, Buffers, Gas chromatography, Polymerase chain reaction, Polymerase chain reaction - quantitative

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Peer-Reviewed Papers
15

References

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