71085

KOD DNA Polymerase

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

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

Properties

• recombinant: expressed in E. coli
• Quality Level: 100
• 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
• UniProt accession no.: P77933
• shipped in: wet ice
• storage temp.: −20°C
• Gene Information: Thermococcus kodakaraensis ... TK_RS00010(3233723)

Description

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

Safety Information

• Pictograms: GHS07
• Signal Word: Warning
• Hazard Statements: H319 - H412
• Precautionary Statements: P264 - P273 - P280 - P305 + P351 + P338 - P337 + P313 - P501
• Hazard Classifications: Aquatic Chronic 3 - Eye Irrit. 2
• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2