TaqDNA
Taq DNA Polymerase Recombinant
Cat. No.
BT15668
Source
Recombinant e.coli contains Thermus aquaticus polymerase gene.
Synonyms
DNA polymerase I thermostable, EC 2.7.7.7, Taq polymerase 1.
Appearance
Purity
Greater than 95.0% as determined by SDS-PAGE.
Usage
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description
Taq DNA Polymerase(a) is a thermostable enzyme of approximately 95 kDa isolated from Thermus aquaticus. This unmodified enzyme replicates DNA at 74°C and exhibits a half-life of 40 minutes at 95°C. The enzyme catalyzes the polymerization of nucleotides into duplex DNA in the 5´~3´ direction in the presence of magnesium and also possesses a 5´~3´ exonuclease activity. Taq DNA Polymerase is recommended for use in PCR but is not recommended for use in DNA sequencing reactions.
Product Specs
Description
Taq DNA Polymerase (a) is a thermostable enzyme with an approximate size of 95 kDa, derived from the bacterium Thermus aquaticus. This enzyme, in its unmodified form, replicates DNA at a temperature of 74 degrees Celsius and demonstrates a half-life of 40 minutes at 95 degrees Celsius. Its function involves catalyzing the polymerization of nucleotides into double-stranded DNA in the 5' to 3' direction, requiring the presence of magnesium. Additionally, it possesses a 5' to 3' exonuclease activity. While recommended for Polymerase Chain Reaction (PCR) applications, Taq DNA Polymerase is not advisable for use in DNA sequencing reactions.
Formulation
Taq DNA Polymerase is provided as a solution containing the following components: 20mM Tris-HCl (pH 8.0), 100mM KCl, 0.1mM EDTA, 1mM DTT, 50% Glycerol, 0.5% NP40, and 0.5% Tween 20.
Unit Definition
One unit of activity is defined as the quantity of enzyme necessary to catalyze the incorporation of 10 nanomoles of deoxynucleotide triphosphates (dNTPs) into acid-insoluble material over a period of 30 minutes at 74 degrees Celsius. This measurement is conducted under specific reaction conditions, including: 50mM Tris-HCl (pH 9.0 at 25 degrees Celsius), 50mM NaCl, 5mM MgCl2, 200 micromolar concentration of each dNTP (dATP, dCTP, dGTP, and a mixture of unlabeled and [3H]dTTP), 10 micrograms of activated calf thymus DNA, and 0.1 mg/ml bovine serum albumin (BSA) in a total reaction volume of 50 microliters.
10x Reaction Buffer With Mgcl2
The 10X Reaction Buffer with MgCl2 consists of: 500mM KCl, 100mM Tris-HCl (pH 9.0 at 25 degrees Celsius), 1% Triton X-100, and 15mM MgCl2. This buffer formulation is optimized for use with a dNTP concentration of 0.2mM for each dNTP.
10x Reaction Buffer Without Mgcl2
The 10X Reaction Buffer without MgCl2 consists of: 500mM KCl, 100mM Tris-HCl (pH 9.0 at 25 degrees Celsius), and 1% Triton X-100. When using this buffer, a separate 25mM MgCl2 Solution should be included in the reaction setup.
Specify Your Own Reaction Conditions
For setting up reactions, choose either Taq DNA Polymerase with the Mg-free 10X Reaction Buffer and a separate 25mM MgCl2 Solution, or Taq DNA Polymerase with the 10X Reaction Buffer containing 15mM MgCl2.
Stability
The enzyme exhibits stability for a period of 5 days when stored at 10 degrees Celsius. For extended storage, it is recommended to store the enzyme at -20 degrees Celsius.
Purity
The purity of the enzyme is determined to be greater than 95.0% as assessed by SDS-PAGE analysis.
Synonyms
DNA polymerase I thermostable, EC 2.7.7.7, Taq polymerase 1.
Source
Recombinant e.coli contains Thermus aquaticus polymerase gene.
Storage Buffer
Compatibility with Reaction Buffers: Taq DNA Polymerase in Storage Buffer. Use of other reaction buffers that do not contain Triton X-100 (final concentration of 0.1%) will result in inactivation of the enzyme.
50mM Tris-HCl (pH 8.0), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 50% glycerol and 1% Triton X-100.
50mM Tris-HCl (pH 8.0), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 50% glycerol and 1% Triton X-100.
Product Science Overview
Discovery and Importance
The discovery of Taq DNA Polymerase revolutionized the field of molecular biology. Before its discovery, DNA polymerases used in PCR were not thermostable, meaning they would denature at the high temperatures required for DNA denaturation during PCR cycles. The thermostability of Taq DNA Polymerase allows it to withstand the high temperatures (up to 95°C) used in PCR, making the process more efficient and reliable .
Recombinant Taq DNA Polymerase
Recombinant Taq DNA Polymerase is produced by cloning the gene encoding the enzyme from Thermus aquaticus and expressing it in Escherichia coli (E. coli). This method allows for large-scale production of the enzyme, ensuring a consistent and high-quality supply for research and industrial applications .
Properties and Function
Taq DNA Polymerase has several key properties that make it ideal for PCR:
- Thermostability: It remains active at high temperatures, with a half-life of more than 40 minutes at 95°C .
- 5’→3’ DNA Polymerase Activity: It synthesizes DNA by adding nucleotides to the 3’ end of a primer annealed to a single-stranded DNA template .
- 5’→3’ Exonuclease Activity: It can degrade DNA from the 5’ end, which is useful for certain applications .
- Deoxynucleotidyl Transferase Activity: It often adds an extra adenine nucleotide to the 3’ end of PCR products, creating “A” overhangs that are useful for cloning .
Applications
Taq DNA Polymerase is widely used in various molecular biology applications, including:
- Standard PCR: Amplification of DNA fragments up to 5 kb in length .
- RT-PCR: Reverse transcription PCR for amplifying RNA sequences by converting them into DNA.
- DNA Sequencing: Sequencing single-stranded DNA templates.
- Genotyping: Identifying genetic variations in organisms.
- Cloning: Creating recombinant DNA molecules for research and industrial purposes .
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