• KOD DNA Polymerase
  • KOD DNA Polymerase

KOD DNA Polymerase

KOD DNA polymerase is derived from a plasmid cloned with the Thermococcus Kodakaraensis DNA polymerase gene and purified by inducing expression in Escherichia coli
$42.00
  • KOD DNA Polymerase

SPECIFICATION

                             KOD DNA Polymerase

KOD DNA polymerase is derived from a plasmid cloned with the Thermococcus Kodakaraensis DNA polymerase gene and purified by inducing expression in Escherichia coli. The super strong 3 '→ 5' exonuclease activity of this enzyme makes its amplification fidelity higher than Pfu DNA Polymerase, with a fidelity about 50 times that of Taq. It also has the characteristic of fast synthesis speed, with a polymerization speed about 5 times that of ordinary Pfu DNA Polymerase and 2 times that of Taq DNA Polymerase, reaching 100-138bp/s. It can obtain high yield amplification products in a short time, especially suitable for high-fidelity amplification of PCR products within 6kb, The amplified DNA is flat ended and can be used for molecular biology experiments such as gene cloning, expression, and mutation analysis.
Under the condition of activity measurement at 75 ℃, the enzyme activity required to consume 10 nmol of dNTPs within 30 minutes to make them acidic insoluble is defined as 1U.
PCR, especially for cloning PCR products and smoothing DNA fragments.
Component 250U 500U
KOD DNA Polymerase (5U/ μ L) 50 μ L 100 μ L
ten × KOD Buffer 1mL 1mL
SuperPure dNTP mix (10mM Each) 100 μ L 200 μ L
Storage: -20 ℃
Suggested PCR conditions (with 50 μ L reaction system as an example)
Component dosage and final concentration
Template Variable < 0.5 μ G
Forward Primer 10 μ M 1 μ L 0.2 μ M
Reverse Primer 10 μ M 1 μ L 0.2 μ M
ten × KOD Buffer 5 μ L 1 ×
DNTP Mixture (10mM each) 1 μ L 0.2mM
KOD DNA Polymerase 0.25-0.5 μ L-
DH2O to 50 μ L-
Attention:
Fill the final volume with sterile PCR grade water to 50 μ L. After calculating the amount of water to be added in practical operation, it is recommended to first add water, then add other ingredients in the above order, and finally add KOD DNA Polymerase. After thoroughly mixing, centrifuge for a few seconds to allow the reaction mixture to settle to the bottom of the tube. Then place the reaction tube in the PCR instrument for amplification.
Mix various components of PCR on an ice bath to prevent KOD DNA polymerase from degrading primers and templates.
High fidelity KOD requires high purity of dNTP, so it is recommended to use the ultra pure dNTP mix matched with this enzyme.
Optimization of PCR conditions
Plasmid or phage template:
The template quantity is 5-20ng, and the cycle parameters are shown in the table below:
Cycling parameters<1kbp target DNA 1-2kbp target
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