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71085

Sigma-Aldrich

KOD DNA Polymerase

High fidelity DNA polymerase designed for accurate PCR amplification of DNA templates for general cloning and cDNA amplification applications.

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Synonym(s):
High fidelity PCR, High fidelity polymerase, KOD polymerase

recombinant

expressed in E. coli

Quality Level

assay

≥90% (homogenous band, SDS-PAGE)

usage

sufficient for 250 reactions

5′ exonuclease activity

<2%, 74 °C (1 h incubation in a reaction with 5′-labeled λ/Sca I digest, per unit enzyme)

secondary activity

nicking (none detected)

feature

Difficult Templates/Specialty Enzymes PCR
High Fidelity PCR
dNTPs included
hotstart: no

manufacturer/tradename

Novagen®

storage condition

OK to freeze

concentration

2.5 unit/μL

technique(s)

PCR: suitable

input

purified DNA

suitability

suitable for PCR

shipped in

wet ice

storage temp.

−20°C

Gene Information

Thermococcus kodakaraensis ... TK_RS00010(3233723)

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vibrant-m

71085

KOD DNA Polymerase

vibrant-m

D8045

AccuTaq LA DNA Polymerase

Gene Information

Thermococcus kodakaraensis ... TK_RS00010(3233723)

Gene Information

-

Gene Information

-

Gene Information

-

assay

≥90% (homogenous band, SDS-PAGE)

assay

-

assay

-

assay

-

recombinant

expressed in E. coli

recombinant

-

recombinant

-

recombinant

-

suitability

suitable for PCR

suitability

suitable for PCR

suitability

suitable for PCR

suitability

suitable for PCR

concentration

2.5 unit/μL

concentration

5 units/μL

concentration

1 unit/μL

concentration

1 unit/μL

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.KOD DNA Polymerase (formerly KOD HiFi DNA Polymerase) is a recombinant form of Thermococcus kodakaraensis KOD1 DNA polymerase. KOD is a high fidelity thermostable DNA polymerase that amplifies target DNA up to 6 kbp with superior accuracy and yield for PCR applications. The enzyme′s 3′→5′ exonuclease-dependent proofreading activity results in a lower PCR mutation frequency than any other commercially available DNA polymerase. The elongation rate and processivity are 5 times and 10 to 15 times higher, respectively, than for Pfu DNA polymerase, resulting in highly accurate and robust yield, in a short reaction time. The enzyme generates blunt-ended PCR products suitable for cloning with the Novagen Perfectly Blunt® and LIC Vector Kits.

Application

KOD DNA Polymerase has been used in high fidelity polymerase chain reaction (PCR) to amplify desired severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) N constructs.

Features and Benefits

  • Higher fidelity than Pfu DNA polymerase excellent for cloning
  • Greater yield extension speed is 2X faster than Taq DNA polymerase and 5X faster than Pfu DNA polymerase
  • Higher processivity sequential nucleotide polymerization is 10- to 15-fold greater than Pfu and Tli DNA polymerases
  • Amplifies plasmid and lambda DNA templates up to 6 kbp
  • Amplifies genomic DNA templates up to 2 kbp
  • No truncated amplification products

Components

  • 250 U KOD DNA Polymerase (2.5 U/μl)
  • 1 ml 10X Buffer #1 for KOD DNA Polymerase (pH 8.0)
  • 1 ml 10X Buffer #2 for KOD DNA Polymerase (pH 8.8)
  • 1 ml 25 mM MgCl
  • 1 ml dNTP 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.

Legal Information

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

pictograms

Exclamation mark

signalword

Warning

Hazard Classifications

Aquatic Chronic 3 - Eye Irrit. 2

Storage Class

10 - Combustible liquids

wgk_germany

WGK 2


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M Nishioka et al.
Journal of biotechnology, 88(2), 141-149 (2001-06-14)
DNA polymerase from Thermococcus kodakaraensis KOD1 (previously Pyrococcus sp. KOD1) is one of the most efficient thermostable PCR enzymes exhibiting higher accuracy and elongation velocity than any other commercially available DNA polymerase [M. Takagi et al. (1997) Appl. Environ. Microbiol.
M Takagi et al.
Applied and environmental microbiology, 63(11), 4504-4510 (1997-11-15)
The DNA polymerase gene from the archaeon Pyrococcus sp. strain KOD1 (KOD DNA polymerase) contains a long open reading frame of 5,013 bases that encodes 1,671 amino acid residues (GenBank accession no. D29671). Similarity analysis revealed that the DNA polymerase
Hirotaka Ata et al.
PLoS genetics, 14(9), e1007652-e1007652 (2018-09-13)
One key problem in precision genome editing is the unpredictable plurality of sequence outcomes at the site of targeted DNA double stranded breaks (DSBs). This is due to the typical activation of the versatile Non-homologous End Joining (NHEJ) pathway. Such
Application of a Spacer-nick Gene-targeting Approach to Repair Disease-causing Mutations with Increased Safety.
Tran, et al.
Bio-protocol, 13, e4661-e4661 (2023)
Despoina A I Mavridou et al.
Current biology : CB, 28(3), 345-355 (2018-02-06)
Animals have evolved a wide diversity of aggressive behavior often based upon the careful monitoring of other individuals. Bacteria are also capable of aggression, with many species using toxins to kill or inhibit their competitors. Like animals, bacteria also have systems

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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