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Sigma-Aldrich

KOD Hot Start Master Mix

ready-to-use solution, 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., 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
NACRES:
NA.55

Quality Level

100

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

storage temp.

−20°C

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

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Yesenia Correa et al.
PloS one, 14(12), e0218837-e0218837 (2019-12-21)
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