SKU: AFG-BOR-102

AffiTaq® DNA Polymerase - With Buffer - With MgCl2 - With dNTP

Vendor AffiGEN
Regular price €600,00 EUR
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AffiTaq® DNA Polymerase - With Buffer - With MgCl2 - With dNTP

Product Overview:

Description: AffiTaq® DNA Polymerase is a high-quality enzyme blend formulated with a proprietary buffer solution and MgCl2 for optimal PCR performance. This variant is tailored specifically for PCR applications, providing flexibility in experimental designs.

Product Compenents: 

Compenents Concentration AFG-BOR-102
AffiTaq® DNA Polymerase 5U/μL 10X (1000U - 200μL)
AffiTaq® Buffer - dNTP free & Mg2+ free 10X 10X (200μL)
AffiTaq® MgCl2 25mM 10X (200μL)
AffiTaq®dNTPs 10mM 10x(800μL)


Product Specifications:

Composition:

  • DNA Polymerase: AffiTaq® sourced from highly purified recombinant enzymes, offering robust and reliable performance.
  • Buffer: A unique proprietary buffer blend designed to optimize enzyme activity and stability during amplification reactions.
  • MgCl2 Concentration: Included at an optimized concentration for enhanced polymerase activity.
  • dNTPs:  serve as the building blocks for DNA synthesis during PCR reactions, providing the necessary nucleotides for polymerase extension.

Storage Conditions:

  • Temperature: Store at -20°C to ensure long-term stability and preservation of enzymatic activity.
  • Stability: This product maintains stable activity for up to 24 months when stored under recommended conditions.

Performance Characteristics:

Key Features:

  • Higher Robustness: AffiTaq® DNA Polymerase ensures high robustness and accuracy during DNA amplification.
  • Enhanced Stability: The proprietary buffer composition and optimized MgCl2 concentration ensure prolonged enzymatic stability.
  • Versatile Usage: Suitable for various PCR applications, including but not limited to, routine PCR, endpoint PCR, and high-throughput PCR.

Applications:

  • PCR Applications: Ideal for routine PCR, genotyping and other molecular biology applications requiring high-quality amplification.
  • Sensitivity and Specificity: Demonstrates exceptional sensitivity and specificity across a wide range of templates and reaction conditions.

Quality Control:

QC Tests Performed:

  • Stringent quality control tests are conducted, including purity assays, activity assays, and specificity assays, to guarantee consistent performance and reliability.

Quality Assurance:

  • Manufactured under ISO-certified conditions with rigorous quality assurance protocols to ensure batch-to-batch consistency and reliability.

Usage Guidelines:

Recommended Protocol:

  • Detailed protocols and guidelines for using AffiTaq® DNA Polymerase are provided, including recommended reaction conditions and cycling parameters for optimal results.
  • Recommended Concentrations: General recommendations for primer and template concentrations are provided for various PCR applications.

Handling Precautions:

  • Follow standard laboratory safety practices when handling enzymes. Avoid repeated freeze-thaw cycles to maintain enzyme activity.

Regulatory Compliance:

Certifications and Compliance:

  • Complies with relevant regulatory standards and is manufactured in accordance with ISO 9001:2015 guidelines.
  • Certified free of contaminants such as endonucleases, exonucleases, and RNases.

Safety Data:

  • Safety data sheets (SDS) detailing hazard warnings, storage, handling, and disposal information are available upon request.

Packaging Information:

Packaging Details:

  • Available in various packaging sizes to accommodate different laboratory requirements, with stringent sealing to prevent contamination.
  • Packaging materials ensure stability and integrity during transportation and storage.

Labeling Information:

  • Each product is labeled with batch numbers, expiration dates, storage conditions, and specific composition details for traceability and quality control purposes.

Suggested protocol for PCR Using AffiTaq® DNA Polymerase - With Buffer - With MgCl2 - With dNTP

Materials Provided:

  1. AffiTaq® DNA Polymerase (5U/μL)
  2. AffiTaq® Buffer (10X)
  3. AffiTaq® MgCl2 (25mM)
  4. AffiTaq®dNTPs (10 mM)

Materials Required but not provided:

  1. Primer pairs specific to the target DNA sequence (25)
  2. Template DNA
  3. PCR tubes/strips and PCR machine
  4. Sterile, nuclease-free water
  5. PCR-grade pipettes and tips

Protocol:

Preparation:

  1. Thaw the AffiTaq® DNA Polymerase set components on ice.
  2. Prepare a master mix containing the following components for each PCR reaction:
    • AffiTaq® DNA Polymerase (5 U/μl)
    • AffiTaq® Buffer (10X)
    • AffiTaq®dNTPs (10 mM)
    • MgCl2 (if not included in the buffer, add as specified in the product instructions)
    • Sterile, nuclease-free water

PCR Reaction Setup:

1. Set up the PCR reaction tubes/strips on ice.

2. Add the following components to each PCR tube:

Compenents Concentration Volume per Reaction Final Concentration
AffiTaq® Buffer - dNTP free & Mg2+ free 10X 5 μL 1X
AffiTaq®dNTPs (10 mM)
10 mM 1 μL 200 μM
AffiTaq® MgCl2 25mM 2 to 8 μL 1 to 4 mM
AffiTaq® DNA Polymerase 5U/μL 0.25 to 0.5 μL 1.25 to 2.5 U
Primer Mix 25 2 μL 1X
Template - - Less than 1μg/Reaction
ddH2O - Adjust to 50 μL -


3. Mix the contents gently by pipetting and briefly spin down the tubes to collect all the liquid at the bottom.

PCR Cycling Conditions:

1. Start the PCR machine and set the following cycling parameters:

  • Initial Denaturation: 95°C for 2-5 minutes (1 cycle)
  • Denaturation: 95°C for 15-30 seconds
  • Annealing: [Optimal temperature]°C for 15-30 seconds
  • Extension: 72°C for [Extension time based on product guidelines] seconds
  • Final Extension: 72°C for 5-10 minutes (1 cycle)
  • Hold at 4-10°C
2. Run the PCR according to the specified cycling parameters for the number of cycles recommended for your specific target.


Post-PCR Steps:

1. Analyze the PCR products using agarose gel electrophoresis or other appropriate methods.

2. Store the remaining PCR products at -20°C for further analysis or downstream applications.

Notes:

  • Use proper aseptic techniques and wear gloves to avoid contamination.
  • Ensure adjustments to annealing temperature and extension time, if necessary, for specific primer-template combinations.
  • Always refer to the specific instructions provided with the AffiTaq® DNA Polymerase set for optimal conditions based on your experiment.