PowerResist Taq Polymerase

Cat. No.: FP1509109

GRADE & PURITY Suitable for molecular biology, EnzymoPure™, for DNA and RNA applications, 5 U/μL

Bioactivity

5 U/μL

Descripción general

  PowerResist Taq Polymerase is a mutant enzyme obtained by site-directed mutagenesis of wild-type Taq enzyme using genetic engineering techniques.
  Compared with the wild-type, this enzyme has higher affinity for templates and stronger resistance to inhibitors. It exhibits high tolerance to endogenous and exogenous interferents in clinical samples, enabling direct amplification of whole blood, crude extracts from fecal samples, and crude extracts from plant samples. This effectively reduces steps such as sample pretreatment and DNA extraction.
  This enzyme has a fast amplification rate, making it suitable for rapid PCR; it also has high detection sensitivity, which is applicable for low-template detection. Additionally, it possesses strong single-base recognition ability and can be used for SNP (Single Nucleotide Polymorphism) detection.
  It can be used in probe-based quantitative real-time PCR (qPCR) detection assays.
  The enzyme is modified with two Taq monoclonal antibodies, which can simultaneously block the 5’-3’ polymerase activity and 5’-3’ exonuclease activity of the Taq enzyme. Before high-temperature heating, the blocking effect of the antibodies on the polymerase activity can effectively inhibit non-specific amplification caused by non-specific annealing of primers or primer dimers under low-temperature conditions, thereby improving the specificity and efficiency of amplification. Meanwhile, the blocking effect on the exonuclease activity can reduce the cleavage of fluorescent probes under low-temperature conditions, ensuring a stable baseline in qPCR and obtaining a good S-shaped amplification curve.
When the amplification reaction system is heated to 95°C for 2.5 minutes, the modified antibodies denature and dissociate, releasing the polymerase activity and exonuclease activity of the Taq enzyme. Therefore, no special inactivation treatment is required, and the enzyme can be used under conventional PCR reaction conditions.
When used in conjunction with the improved amplification buffer, the enzyme can be activated quickly, effectively increasing the amount of reaction products and enhancing the sensitivity, specificity, and anti-interference ability of the PCR reaction.

Component

FP1509109	Component	250U	1KU	2.5KU	4×2.5KU	Storage
FP1509109A	PowerResist Taq Polymerase (5U/μL)	50μl	200μl	500μl	4×500μl	-20℃. Avoid freeze/thaw cycle.
FP1509109B	5×GN Buffer（Without Mg²⁺）	1ml	4×1ml	10×1ml	40×1ml	-20℃. Avoid freeze/thaw cycle.
FP1509109C	100 mM Mg²⁺	100μl	400μl	1ml	4×1ml	-20℃. Avoid freeze/thaw cycle.

Scope of ApplicationSuitable for hot-start qPCR amplification, direct whole blood amplification, anti-interference amplification, rapid PCR, and ARMS (Amplification Refractory Mutation System).

Precautions

1. For most amplification reactions, the initial denaturation time can be set at 95°C for 2.5 minutes. If the template is complex, the denaturation time can be adjusted appropriately.
2. The extension time should be determined according to the length of the PCR product. For example, when amplifying a 2kb fragment of human genomic DNA, the extension time can be as fast as 15 seconds. If it is a complex template (such as using whole blood as a template for amplification), the extension time can be appropriately prolonged.

Other Precautions

1. When performing PCR amplification using whole blood as a template, the volume of whole blood added is recommended to be controlled within 20% (v/v) of the reaction system volume.
2. When used for fluorescent quantitative PCR, the volume of whole blood added is recommended to be ≤ 5% (v/v) of the reaction system volume. Excessive addition of blood will affect the collection of fluorescent signals.

Usage Method

Preparation before Reaction System Configuration

1.1 Dissolve and mix various solutions required for the reaction at room temperature or 4°C, and then place them in an ice bath or ice box. It is recommended to aliquot the reaction liquids for use to avoid repeated freezing and thawing.
1.2 The reaction system volume can be adjusted in proportion according to the experimental project requirements to ensure that the final concentration of each component is consistent.
1.3 Refer to the following table to set up the fluorescent quantitative PCR reaction system. It is recommended to complete the system configuration on an ice bath or ice box.

Component	Volume Added	Final Concentration
5×GN Buffer (without Mg²⁺)	10 μL	1×
dNTPs (25 mM each)	0.4 μL	0.2 mM
Template DNA	5 - 10 μL	—
Forward Primer (10 μM)	1 μL	0.2 μM
Reverse Primer (10 μM)	1 μL	0.2 μM
Probe (10 μM)	0.5 μL	0.1 μM
PowerResist Taq Polymerase (5 U/μL)	0.5 μL	2.5 U/50 μL
MgCl₂ (100 mM)	1 μL	2 mM
ddH₂O	Make up to 50 μL	—
Total Volume	50 μL	—

*Recommended amounts of different types of templates in a 50 μL reaction volume are as follows:
Mammalian genomic DNA: 0.1 - 1 μg
E. coli genomic DNA: 10 - 100 ng
Plasmid DNA: 0.1 - 10 ng
1.4 The recommended amount of Mg²⁺ is 2 mM - 6 mM.
1.5 Avoid vigorous shaking during preparation. It can be mixed by gently pipetting or slightly vortexing, and then centrifuged at room temperature for a few seconds.

Reaction Procedure

1. Conventional Qualitative PCR (taking the amplification of a 1 kb fragment as an example)

Step Name	Temperature	Time	Number of Cycles
Heat Shock	95°C	2min30s	1
Denaturation	95°C	20s	30 - 35
Annealing	55°C	20s	30 - 35
Extension	72°C	40s	30 - 35
Extension	72°C	7min	1
Storage	12°C	∞ (Indefinite)	1

2. Fluorescent Quantitative PCR

Step Name	Temperature	Time	Number of Cycles
Heat Shock	95°C	2min30s	1
Denaturation	95°C	15s	35 - 40
Annealing + Extension	55°C	40s	35 - 40

a. PCR reaction conditions (including temperature, time, number of cycles, etc.) need to be flexibly adjusted according to factors such as templates, primers, PCR product length, and GC content.
b. The extension time should be set according to the length of the PCR product. Usually, the fastest extension time for each kb product is 15 seconds. For example: when the length of the PCR product is 1kb, the extension time can be set to 15 seconds; when the product length is 2kb, the extension time can be set to 30 seconds; if the amplification effect is not good, it can be appropriately extended to 45 seconds, and so on for fragments of other lengths.

Specifications

Product Name

PowerResist Taq Polymerase

Grado

for DNA and RNA applications, Suitable for molecular biology, EnzymoPure™

Especificaciones y pureza

Suitable for molecular biology, EnzymoPure™, for DNA and RNA applications, 5 U/μL

Bioactividad

5 U/μL

Tipo de molécula

Enzyme

Almacenamiento y envío

Concentración

5 U/μL

Condiciones de almacenamiento de almacenamiento

Store at -20°C,Avoid repeated freezing and thawing

Enviado en

Ice chest + Ice pads

Estabilidad y almacenamiento

Store at -20℃ long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Definición de unidad

One unit (U) of activity is defined as the activity that catalyzes the incorporation of 10 nanomoles (nmol) of total nucleotides into acid-insoluble substances within 30 minutes at 74°C, using activated salmon sperm DNA as the template/primer.