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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

Synonym(s):

High fidelity master mix, Hot start PCR, Hot start master mix

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About This Item

UNSPSC Code:
41105500

Quality Level

form

ready-to-use solution

usage

sufficient for 100 reactions
sufficient for 500 reactions

feature

Difficult Templates/Specialty Enzymes PCR
High Fidelity PCR
dNTPs included
hotstart

manufacturer/tradename

Novagen®

storage condition

OK to freeze

technique(s)

PCR: suitable (2X PCR Master Mix for high fidelity)
PCR: suitable

input

purified DNA

shipped in

dry ice
wet ice

storage temp.

−20°C

Related Categories

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.
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. The mix contains KOD Hot Start DNA Polymerase, two monoclonal antibodies, ultra-pure deoxynucleotides, and reaction buffer with MgSO4. The monoclonal antibodies inhibit the DNA polymerase and 3′→5′ exonuclease activities at ambient temperatures.

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.

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 mL KOD Hot Start Master Mix

References:
Kitabayashi, M., et al. 2002. Biosci. Biotechnol. Biochem.66 (10), 2194. Fujii, S., et al. 1999. J. Mol.

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

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Kim A Lagerborg et al.
bioRxiv : the preprint server for biology (2021-03-25)
The rapid global spread and continued evolution of SARS-CoV-2 has highlighted an unprecedented need for viral genomic surveillance and clinical viral sequencing. Amplicon-based sequencing methods provide a sensitive, low-cost and rapid approach but suffer a high potential for contamination, which
Kim A Lagerborg et al.
Nature microbiology, 7(1), 108-119 (2021-12-16)
The global spread and continued evolution of SARS-CoV-2 has driven an unprecedented surge in viral genomic surveillance. Amplicon-based sequencing methods provide a sensitive, low-cost and rapid approach but suffer a high potential for contamination, which can undermine laboratory processes and
Kothai Parthiban et al.
mAbs, 11(5), 884-898 (2019-05-21)
The construction of large libraries in mammalian cells allows the direct screening of millions of molecular variants for binding properties in a cell type relevant for screening or production. We have created mammalian cell libraries of up to 10 million
Yesenia Correa et al.
PloS one, 14(12), e0218837-e0218837 (2019-12-21)
The gut microbiota of insects is composed of a wide range of microorganisms which produce bioactive compounds that protect their host from pathogenic attack. In the present study, we isolate and identify the fungus Chrysosporium multifidum from the gut of
Fantao Kong et al.
Bio-protocol, 8(3), e2718-e2718 (2018-02-05)
Chlamydomonas reinhardtii is frequently used as a model organism to study fundamental processes in photosynthesis, metabolism, and flagellar biology. Versatile tool boxes have been developed for this alga ( Fuhrmann et al., 1999 ; Schroda et al., 2000 ; Schroda

Articles

A PCR master mix is a batch of PCR or RT-PCR reagents that can be divided among many PCR reaction tubes. It usually includes DNA polymerase, dNTPs, MgCl2 and buffer. Make your own master mix or choose a commercial one.

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.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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