SKU: AFG-BOR-100

AffiTaq® DNA Polymerase - With Buffer - With MgCl2 - Without dNTP

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AffiTaq® DNA Polymerase - With Buffer - With MgCl2 - Without 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 where the addition of dNTPs is done separately, providing flexibility in experimental designs.

Product Compenents: 

Compenents Concentration AFG-BOR-100 AFG-BOR-101
AffiTaq® DNA Polymerase 5U/μL 1X (200U - 40μL) 10X (1000U - 200μL)
AffiTaq® Buffer - dNTP free & Mg2+ free 10X 1X (40μL) 10X (200μL)
AffiTaq® MgCl2 25mM 1X (40μL) 10X (200μL)



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.

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 - Without dNTP

Materials Provided:

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

Materials Required but not provided:

  1. Primer pairs specific to the target DNA sequence (25)
  2. Template DNA
  3. Nucleotides (dNTPs) (10 mM)
  4. PCR tubes/strips and PCR machine
  5. Sterile, nuclease-free water
  6. 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)
    • MgCl2 (if not included in the buffer, add as specified in the product instructions)
    • Sterile, nuclease-free water
    • Note: Exclude dNTPs from the master mix as they will be added separately.

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
dNTP 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.