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Taq DNA Polymerase, Recombinant


Highly thermostable DNA polymerase for routine PCR.
  
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PCR using <em>Taq</em> DNA Polymerase in 10X <em>Taq</em> Buffer with (NH4)<sub>2</sub>SO<sub>4</sub> and different MgCl<sub>2</sub> concentrations

A 950 bp single copy gene from human genomic DNA was amplified at a wide range of Mg2+ concentrations.

M – GeneRuler 100 bp Plus DNA Ladder

Thermo Scientific Taq DNA Polymerase is a highly thermostable DNA polymerase from the thermophilic bacterium Thermus aquaticus. The enzyme catalyzes 5’→3’ synthesis of DNA, has no detectable 3’→5’ exonuclease (proofreading) activity and possesses low 5’→3’ exonuclease activity. In addition, Taq DNA Polymerase exhibits deoxynucleotidyl transferase activity, which frequently results in the addition of extra adenines at the 3’-end of PCR products. Recombinant Taq DNA Polymerase is the ideal tool for standard PCR of templates 5 kb or shorter.

PCR Master Mix is a 2X concentrated solution of Taq DNA Polymerase, dNTPs, and all of the components required for PCR, except DNA template and primers. This pre-mixed formulation saves time and reduces contamination due to a reduced number of pipetting steps required for PCR set up. The mix is optimized for efficient and reproducible PCR.

Highlights

  • Thermostable – half life is more than 40 min at 95°C
  • Generates PCR products with 3’-dA overhangs
  • Supplied with two buffers – 10X Taq Buffer with KCl and 10X Taq Buffer with (NH4)2SO4. The latter allows for PCR at wide range of magnesium concentrations and decreases unspecific priming
  • Incorporates modified nucleotides (e.g., biotin-, digoxigenin-, fluorescently-labeled nucleotides)

Applications

  • Routine PCR amplification of DNA fragments up to 5 kb
  • High throughput PCR
  • DNA labeling (see Reference 1,2,3)

Includes

Taq DNA Polymerase (recombinant):

  • Taq DNA Polymerase 5 U/µL
  • 10X Taq Buffer with KCl
  • 10X Taq Buffer with (NH4)2SO4
  • 25 mM MgCl2

Taq DNA Polymerase (recombinant), LC:

  • Taq DNA Polymerase 1 U/µL
  • 10X Taq Buffer with KCl
  • 10X Taq Buffer with (NH4)2SO4
  • 25 mM MgCl2

PCR Master Mix (2X):

  • Taq DNA polymerase (0.05 U/µL), reaction buffer, 4 mM MgCl2, and 0.4 mM of each dNTP
  • Nuclease-free water

Note

  • The error rate of Taq DNA Polymerase in PCR is 2.2 x 10-5 errors per nt per cycle, as determined by a modified method described in (see Reference 4). Accordingly, the accuracy of PCR is 4.5 x 104. Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs.
  • The 10X Taq Buffer without Detergent is recommended for microarray experiments.
  
Storage Condition-20 C
HazardousNo
10X Taq Buffer with (NH4)2SO4750 mM Tris-HCl (pH 8.8 at 25°C), 200 mM (NH4)2SO4, 0.1% (v/v) Tween 20.
10X Taq Buffer with KCl100 mM Tris-HCl (pH 8.8 at 25°C), 500 mM KCl, 0.8% (v/v) Nonidet P40.
Definition of Activity Unit
  • One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 min at 70°C.
  • Enzyme activity is assayed in the following mixture: 67 mM Tris-HCl(pH 8.8 at 25°C), 6.7 mM MgCl2, 1 mM 2-mercaptoethanol, 50 mM NaCl, 0.1 mg/mL BSA, 0.75 mM activated calf thymus DNA, 0.2 mM of each dNTP, 0.4 MBq/mL [3H]-dTTP.
InactivationInactivated by phenol/chloroform extraction.
InhibitionInhibitors: ionic detergents (deoxycholate, sarkosyl and SDS) at concentrations higher than 0.06, 0.02 and 0.01%, respectively (5).
Molecular Weight94 kDa monomer.
Quality Control
  • The absence of endo-, exodeoxyribonucleases, and ribonucleases confirmed by appropriate quality tests.
  • Functionally tested in PCR.
SourceE. coli cells with a cloned pol gene from Thermus aquaticus YT1.
Storage BufferThe enzyme is supplied in 20 mM Tris-HCl (pH 8.0), 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.5% (v/v) Nonidet P40, 0.5% (v/v) Tween 20, and 50% (v/v) glycerol.

References

  1. D. Celeda et al., PCR amplification and simultaneous digoxigenin incorporation of long DNA probes for fluorescence in situ hybridization. BioTechniques. 12, 98-102 (1992).

  2. U. Finckh et al., Producing single-stranded DNA probes with the Taq DNA polymerase: a high yield protocol. BioTechniques. 10, 35-39 (1991).

  3. H. Yu et al., Cyanine dye dUTP analogs for enzymatic labeling of DNA probes. Nucleic Acids Res. 22, 3226-3232 (1994).

  4. K. S. Lundberg et al., High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus. Gene. 108, 1-6 (1991).

  5. R. S. Weyant et al., Effect of ionic and nonionic detergents on the Taq polymerase. BioTechniques. 9, 308-309 (1990).

MSDS & Certificates

pdf   MSDS (US) Polymerase
pdf   MSDS (US) - 2X PCR Master Mix (K1071,K1072)
pdf   MSDS (JP) - Taq DNA Polymerase (recombinant)
pdf   MSDS (JP) - Taq DNA Polymerase (recombinant), 10X Taq Buffer with KCl

Protocols and Product Manuals

pdf   EP0401 Recombinant Taq DNA Polymerase - Product Information
pdf   EP0402 Recombinant Taq DNA Polymerase - Product Information
pdf   EP0403 Recombinant Taq DNA Polymerase - Product Information
pdf   EP0404 Recombinant Taq DNA Polymerase - Product Information
pdf   EP0405 Recombinant Taq DNA Polymerase - Product Information
pdf   EP0406 Recombinant Taq DNA Polymerase - Product Information
pdf   K-1071 PCR Master Mix (2X) - Product Information
pdf   K-1072 PCR Master Mix (2X) - Product Information
pdf   Analysis of Recombinant Clones
pdf   Components of the Reaction Mixture
pdf   Cycling Parameters Protocol
pdf   Guidelines for Preventing Contamination of PCR Protocol
pdf   Guidelines for Primer Design Protocol
pdf   Standard PCR (reaction set up) Protocol

Product Guides and Selectors

pdf   PCR Reagents Selection Guide