Taq Plus DNA
Taq Plus DNA Polymerase Recombinant
Cat. No.
BT15602
Source
Recombinant E.coli contains Thermus aquaticus polymerase gene.
Synonyms
Taq Plus DNA Polymerase, Taq Plus DNA, TaqPDNA.
Appearance
Purity
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 Plus is a mixture of Taq and Pfu. Taq Plus, which is used to improve the reliability and yield of conventional primer extension reaction. Taq Plus has two following advantages over Taq: (1) high fidelity with an error frequency 1.6/106 (or 0.0016/103) during DNA synthesis. (2) Taq Plus increases the efficiency of polymerization reaction, resulting in a great percentage of extenuation reaction completion up to 10 kb to 30 kb. Pfu has a temperature optimum between 72-78°C and remains > 95% active following 1-hour incubation at 95°C.
Product Specs
Description
Taq Plus, a blend of Taq and Pfu DNA polymerases, enhances the fidelity and yield of standard PCR reactions. Its advantages over Taq polymerase include a higher fidelity rate of 1.6/10^6 (or 0.0016/10^3) during DNA synthesis and increased polymerization efficiency, leading to a greater percentage of successful extensions, even for fragments up to 10 kb to 30 kb. Pfu polymerase, with an optimal temperature range of 72-78°C, retains over 95% activity after 1 hour at 95°C.
Unit Definition
One unit of Taq Plus catalyzes the incorporation of 10 nanomoles of deoxyribonucleotides into a polynucleotide strand over 30 minutes at 74°C.
Formulation
5 units per microliter (5 U/µl)
Note
To ensure optimal activity, combine all reagents, including Taq Plus, immediately prior to use.
Reaction Conditions
DNA synthesis is carried out in a 100 µl reaction volume containing 20-200 µM dNTPs, 0.3-1 µM primers, 0.1-0.250 mg template DNA, 10 µl of 10X reaction buffer, and 2.5-5 units of Taq Plus. After gently mixing the reaction components, briefly centrifuge the tube and add a layer of light mineral oil. Initial denaturation is performed at 95°C for 5 minutes, followed by annealing at 40-68°C for 5 minutes to allow primer binding to the template DNA.
Optimization Of Dna Synthesis
For optimal results, add the reaction components in the following sequence:
1. H2O
2. 10X reaction buffer
3. dNTPs
4. DNA template and primers
5. Taq Plus
Stability
Taq Plus is stable for 5 days when stored at 10°C. For prolonged storage, store at -20°C.
Synonyms
Taq Plus DNA Polymerase, Taq Plus DNA, TaqPDNA.
Source
Recombinant E.coli contains Thermus aquaticus polymerase gene.
Storage Buffer
20mM Tris-HCl (pH 8.0), 1mM DTT, 0.1mM EDTA, 100mM KCl, Stabilizers, and 50%glycerol.
10x Reaction Buffer
200 mM TrisHCI (pH 8.8)
100 mM KCI
100 mM (NH4)2SO4
20 mM Mg SO4
1% Triton X-100
1 mg/ml bovine serum albumin (BSA).
100 mM KCI
100 mM (NH4)2SO4
20 mM Mg SO4
1% Triton X-100
1 mg/ml bovine serum albumin (BSA).
Product Science Overview
Origin and Discovery
The original Taq DNA polymerase was first isolated from Thermus aquaticus, a bacterium that thrives in hot springs and hydrothermal vents. The enzyme’s ability to remain stable and active at high temperatures made it an ideal candidate for PCR, a revolutionary technique developed by Kary Mullis in the 1980s. The recombinant version of Taq DNA polymerase is produced by cloning the gene encoding the enzyme into Escherichia coli (E. coli), allowing for large-scale production and purification .
Characteristics and Properties
Taq Plus DNA Polymerase Recombinant possesses several key characteristics that make it suitable for PCR applications:
- Thermostability: The enzyme remains active at temperatures up to 95°C, which is essential for the denaturation step in PCR.
- 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: This activity allows the enzyme to degrade the RNA strand of RNA-DNA hybrids, which is useful in certain applications .
- Deoxynucleotidyl Transferase Activity: Taq DNA polymerase often adds an extra adenine nucleotide to the 3’ end of PCR products, which can be useful for cloning purposes .
Applications
Taq Plus DNA Polymerase Recombinant is used in various molecular biology applications, including:
- Standard PCR: Amplification of DNA fragments up to 5 kilobases (kb) in length.
- High-Throughput PCR: Suitable for large-scale PCR reactions.
- DNA Labeling: Incorporation of modified nucleotides for labeling DNA .
- RT-PCR: Reverse transcription PCR for amplifying RNA targets.
- Sequencing: Used in cycle sequencing and other sequencing techniques .
Advantages of Recombinant Production
Producing Taq DNA polymerase recombinantly in E. coli offers several advantages:
- Consistency: Recombinant production ensures a consistent and reliable supply of the enzyme.
- Purity: The enzyme can be purified to a high degree, reducing the risk of contaminants that could interfere with PCR.
- Cost-Effectiveness: Large-scale production in E. coli is more cost-effective compared to isolating the enzyme from natural sources .
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