HiFi DNA Polymerase
Description

HiFi DNA polymerase is ideal for amplification of DNA fragments that require high-fidelity. High fidelity is achieved by an optimal blend of high performance YEAtaq DNA polymerase and Accu DNA polymerase, a Pyrococcus proofreading (3'→5' exonuclease activity ) enzyme. This formulation achieves greater yields with higher fidelity than standard DNA polymerases such as Taq. HiFi is a quality choice for sensitive applications. Because of the presence of Accu DNA polymerase in the blend, dUTP, dITP and primers containing these nucleotides should not be used in PCR because they hinder DNA synthesis

HiFi DNA polymerase is offered in two different packages, one with dNTPs mix and the other without. HiFi DNA Polymerase is also available in 2× ready-to-use premix format with a green dye.

Features

  • Robust amplification with minimal optimization.
  • High yields of PCR products.
  • Higher sensitivity compared to conventional Taq DNA Polymerase.
  • Amplification of long targets up to 6 kb from genomic DNA
  • Generates 3'-dA overhangs.
  • Incorporates modified nucleotides.
  • The error rate of HiFi DNA Polymerase in PCR is 8.3×10-6 errors per nucleotide per cycle.
Applications

  • Cloning and expression
  • Blunt end amplification products
  • Mutation analysis
  • Efficient & highly sensitive PCR use
  • Routine PCR amplification of DNA fragments up to 6 kb from genomic DNA
  • RT-PCR
Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 72

Quality Control

  • Nuclease activity is not detected after incubation of 1 μg λ /Hind III DNA with 5 units of HiFi DNA polymerase in 50 μl reaction buffer for 18 hours at 37 °C.
  • The absence of endo-, exodeoxyribonucleases and ribonucleases is confirmed by appropriate tests. Functional test is performed by PCR.
Patterns/Disclaimer

Some applications in which this product can be used may be covered by patents issued and applicable in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a patent license depending upon the particular application and country in which the product is used.

Results

Specification
Store at

 

# FYT003-500U (500 units)
HiFi DNA Polymerase 100 μl (5 U/μl)
10× Reaction Buffer 2 ml
dNTPs Mix 200 μl (10 mM)
# FYT033-500U (500 units)
HiFi DNA Polymerase 100 μl (5 U/μl)
10× Reaction Buffer 2 ml
# FYT301-100P (100 preps)
HiFi DNA Polymerase Premix (2×) with green dye 1.25 ml x 2

 

Storage Buffer

50 mM Tris-HCl (pH 9.0), 100 mM NaCl, 0.1 mM EDTA, 1% Triton X-100, 5 mM DTT, 50% Glycerol, Stabilizers

10× Reaction Buffer*
100 mM KCl, 20 mM MgSO4·7H2O, 200 mM Tris-HCl (pH 8.8), 1% Triton X-100, 100 mM (NH4)2SO4, 1 mg/ml BSA
 
* The reaction buffer is supplied as a 10× concentrate and should be diluted before use.
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