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D4545

Sigma-Aldrich

Taq DNA Polymerase from Thermus aquaticus

with 10× PCR reaction buffer without MgCl2

CAS Number:
Enzyme Commission number:
MDL number:

Quality Level

recombinant

expressed in E. coli

form

liquid

usage

sufficient for 1500 reactions
sufficient for 250 reactions
sufficient for 50 reactions
sufficient for 5000 reactions

feature

dNTPs included: no
hotstart: no

concentration

5 units/μL

technique(s)

PCR: suitable

color

colorless

input

purified DNA

suitability

suitable for PCR and automated sequencing reactions

application(s)

agriculture

shipped in

wet ice

storage temp.

−20°C

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General description

Taq polymerase is a thermostable DNA polymerase named after the thermophilic bacterium Thermus aquaticus. A stable deoxyribonucleic acid (DNA) polymerase (with a temperature optimum of 80°C) has been purified from the extreme thermophile Thermus aquaticus. The enzyme is free from phosphomonoesterase, phosphodiesterase and single-stranded exonuclease activities. Maximal activity of the enzyme needs all four deoxyribonucleotides and activated calf thymus DNA.

Application

Taq DNA Polymerase from Thermus aquaticus has been used:
  • in the process of DNA extraction (during gene amplification and sequencing)
  • in genotyping
  • in polymerase chain reaction (PCR) to study the constitutive production of epithelial neutrophil activating peptide 78 (ENA-78) and interleukin-8 (IL-8)
  • for amplification of RNA from primary endothelial cells by conventional PCR

Biochem/physiol Actions

Taq polymerase catalyzes oligonucleotide primer-driven, DNA template dependent incorporation of dNTPs into complimentary DNA strands. It displays both 5′ to 3′ polymerase and exonuclease activities.

Features and Benefits

  • MgCl2 provided in a separate tube to allow MgCl2 optimization
  • Can withstand repeated heating to 95 °C without significant loss of activity

Packaging

Taq DNA Polymerase with 10× reaction buffer without MgCl2. Includes a separate tube of 25 mM MgCl2
Taq DNA polymerase comes with the choice of an optimized 10× reaction buffer including MgCl2 (D1806) or a 10× reaction buffer without MgCl2 plus a separate tube of MgCl2 for titration (D4545). The latter option may be necessary to determine optimal conditions for amplification.

Other Notes

Taq DNA Polymerase is a specialized thermostable enzyme isolated from the thermophilic bacterium Thermus aquaticus. The recombinant form of this enzyme is expressed in E. coli. This 94 kDa protein shows no detectable levels of contaminating endonucleases or exonucleases by SDS-PAGE. It has both 5′→3′ polymerase and exonuclease activity.

Unit Definition

One unit will incorporate 10 nmol of total dNTPs into acid-precipitable DNA in 30 min at 74 °C.

Legal Information

Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: US 8,404,464 and US 7,972,828. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

A Chien et al.
Journal of bacteriology, 127(3), 1550-1557 (1976-09-01)
A stable deoxyribonucleic acid (DNA) polymerase (EC 2.7.7.7) with a temperature optimum of 80 degrees C has been purified from the extreme thermophile Thermus aquaticus. The enzyme is free from phosphomonoesterase, phosphodiesterase and single-stranded exonuclease activities. Maximal activity of the
Nick A Bersinger et al.
Fertility and sterility, 89(5 Suppl), 1530-1536 (2007-09-01)
To analyze the constitutive production of epithelial neutrophil activating peptide 78 (ENA-78) and interleukin-8 (IL-8) by epithelial cells and the response of these cells to cytokine stimulation. In vitro study using eutopic endometrial tissue. University hospital. Cycling women undergoing laparoscopy
Anna Bobrowska et al.
PloS one, 6(6), e20696-e20696 (2011-06-17)
Huntington's disease (HD) is a progressive neurodegenerative disorder for which there is no effective disease modifying treatment. Following-on from studies in HD animal models, histone deacetylase (HDAC) inhibition has emerged as an attractive therapeutic option. In parallel, several reports have
Ryan E Davey et al.
Stem cells (Dayton, Ohio), 24(11), 2538-2548 (2006-07-11)
Highly ordered aggregates of cells, or niches, regulate stem cell fate. Specific tissue location need not be an obligatory requirement for a stem cell niche, particularly during embryogenesis, where cells exist in a dynamic environment. We investigated autoregulatory fixed-location-independent processes
Rita Horvath et al.
Brain : a journal of neurology, 129(Pt 7), 1674-1684 (2006-04-20)
Mutations in the gene coding for the catalytic subunit of the mitochondrial DNA (mtDNA) polymerase gamma (POLG1) have recently been described in patients with diverse clinical presentations, revealing a complex relationship between genotype and phenotype in patients and their families.

Articles

Introduction and Historical Timelines

Learn about the history of the polymerase chain reaction (PCR), from the basic principles that proceeded its discovery to the awarding of a Nobel Prize for Chemistry and more recent developments such as real-time PCR (qPCR) and digital PCR.

Polymerase Chain Reaction

The polymerase chain reaction is one of the most widely used techniques in molecular biology. The PCR process consists of three main steps, Denaturation, Annealing & Extension

Protocols

Hot Start dNTP protocol to reduce non-specific amplification

Protocol using hot start dNTPs. Method includes modified nucleoside triphosphates that block DNA polymerase nucleotide incorporation during hot start PCR to increase specificity. Compatible with a variety of PCR reagents.

dNTP Mediated Hot Start PCR Protocol

Hot Start dNTPs are modified with a thermolabile protecting group at the 3’ terminus. The presence of this modification blocks nucleotide incorporation by DNA polymerase until the nucleotide protecting group is removed during a heat activation step.

Standard PCR Protocol

Learn standard PCR protocol steps and review reagent lists or cycling parameters. This method for routine PCR amplification of DNA uses standard Taq DNA polymerase.

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