Thermo Scientific™ Phusion Blood Direct PCR Kit

The Thermo Scientific™ Phusion Blood Direct PCR Kit is a high-performance solution for amplifying DNA directly from whole blood without the need for a separate DNA purification step. Leveraging the modified Phusion Hot Start II High-Fidelity DNA Polymerase, this kit ensures robust amplification, exceptional accuracy, and high tolerance to PCR inhibitors typically found in blood.

Key Features:

1. Direct PCR from Whole Blood:
   • Eliminates the need for DNA extraction, saving time and resources.
2. High Resistance to PCR Inhibitors:
   • Effective with samples containing up to 40% whole blood.
3. High Fidelity and Accuracy:
   • The Phusion Hot Start II DNA Polymerase provides exceptional proofreading capability, making it ideal for demanding applications like cloning.
4. Universal Application:
   • Compatible with blood samples from various species, different anticoagulants (e.g., EDTA, heparin), and storage conditions.
5. Fast and Efficient Protocol:
   • Minimal hands-on time with extremely short PCR cycles for rapid results.

Specifications:

Applications:

1. Genotyping:
   • Enables fast and accurate genotyping directly from whole blood samples.
2. Cloning:
   • High-fidelity polymerase ensures accurate amplification for downstream cloning applications.
3. Forensic Analysis:
   • Direct PCR from trace blood samples simplifies workflows and reduces processing time.
4. Pathogen Detection:
   • Amplifies target DNA from blood samples in infectious disease studies.
5. Veterinary and Comparative Genomics:
   • Analyzes blood samples from various species for genetic studies.

Kit Contents:

Reaction Protocol:

1. Setup:
   • Prepare the PCR mix using the provided components.
   • Add up to 40% whole blood directly to the reaction mix.
2. Thermal Cycling:
   • Follow a fast PCR protocol optimized for the Phusion Blood II DNA Polymerase:
     • Initial Denaturation: 98°C for 30 seconds.
     • Denaturation: 98°C for 5–10 seconds.
     • Annealing: Use a temperature optimized for the primers (generally higher than for Taq polymerases).
     • Extension: 72°C for 15–30 seconds/kb.
     • Final Extension: 72°C for 5 minutes.
3. Analysis:
   • Analyze PCR products using gel electrophoresis or other detection methods.

Advantages:

1. Time and Cost Savings:
   • Direct PCR eliminates the need for time-consuming and expensive DNA extraction steps.
2. High Yield and Specificity:
   • Robust amplification even with complex or inhibitor-rich blood samples.
3. Versatility:
   • Compatible with a wide range of species, anticoagulants, and storage conditions.
4. Convenience:
   • Pre-formulated buffer with dNTPs and MgCl₂ simplifies reaction setup.

Storage and Handling:

• Store all components at -20°C. Avoid repeated freeze-thaw cycles to maintain polymerase activity.
• Handle blood samples in accordance with biosafety guidelines.

Product Details:

The Thermo Scientific™ Phusion Blood Direct PCR Kit is an ideal solution for researchers requiring efficient and high-fidelity amplification directly from whole blood. Its ease of use, speed, and exceptional performance make it indispensable for applications in genomics, diagnostics, and molecular biology research.