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:
- AffiTaq® DNA Polymerase (5U/μL)
- AffiTaq® Buffer (10X)
- AffiTaq® MgCl2 (25mM)
- AffiTaq®dNTPs (10 mM)
Materials Required but not provided:
- Primer pairs specific to the target DNA sequence (25)
- Template DNA
- PCR tubes/strips and PCR machine
- Sterile, nuclease-free water
- PCR-grade pipettes and tips
Protocol:
Preparation:
- Thaw the AffiTaq® DNA Polymerase set components on ice.
- 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
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.