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Direct PCR—Phire and Phusion Kits & Master Mixes
The Thermo Scientific Direct PCR approach delivers unprecedented convenience for DNA amplification by allowing PCR directly from unpurified samples. Significant savings in time and cost can be achieved in applications such as plant and animal genotyping. Learn more about the Direct PCR method.
How does Thermo Scientific direct PCR work?
Thermo Scientific direct PCR solutions are based on the engineered Thermo Scientific Phire and Phusion DNA polymerases, which allow highly robust amplification and tolerance to many PCR inhibitors present in unpurified samples. Both DNA polymerases are fused to a small DNA-binding protein, which helps achieve high tolerance to inhibitors, increased speed, and better yields.
Key features
- Optimized formats for animal and human tissues, plant materials, and blood samples
- Direct protocol for quick sample-to-PCR workflow
- Dilution and storage protocol for flexible sample handling
- Sample storage in dilution buffer for up to four weeks before using in PCR
Customer stories on Phire and Phusion direct PCR approach
Two short protocols to suit your needs
Figure 1. Direct and dilution protocols for a versatile and efficient workflow. (A) Direct protocol offers minimal hands-on time. (B) Dilution protocol offers workflow flexibility and optimization.
Thermo Scientific direct PCR master mixes include electrophoresis tracking dyes and a density reagent, which allows direct loading of PCR products on gels, thereby simplifying direct PCR workflows.
Perform PCR directly on an unpurified sample in under 30 min
How to perform the direct vs. dilution PCR protocols
Master mixes and kits available based on sample types
Phire Tissue Direct PCR Master Mix is designed for DNA amplification from animal and human tissues. The master mix contains Phire Hot Start II DNA Polymerase and is specially formulated for performing PCR in the presence of different tissue-derived inhibitors such as collagen, melanin and eumelanin (hair, skin), or myoglobin (muscle). The master mix is supplied with Thermo Scientific DNARelease Additive, which can be used to improve the release of DNA from difficult tissues.
Figure 2. DNA fragments from different human and animal tissue samples were amplified with Phire Tissue Direct PCR Master Mix. Mouse (1) tail, (2) ear, and (3) hair; bird (4) muscle and (5) feather; human (6) hair, (7) tooth, (8) nail, and (9) saliva; (10) zebrafish muscle; and (11) fruit fly.
M: Thermo Scientific O’GeneRuler Express DNA Ladder, +: Positive control reaction, –: No-template control
Phire Plant Direct PCR Master Mix is designed to amplify DNA directly from plant samples. The master mix contains Phire Hot Start II DNA Polymerase and is specially formulated for performing PCR from samples with plant-derived inhibitors such as complex polysaccharides and polyphenols.
Figure 3. Amplification of a 0.5 kb DNA fragment directly from leaves and seeds from different plants. A 0.5 mm punch of leaf (L) or a small piece of crushed seed (S) was placed directly into 50 μL of Phire Plant Direct PCR Master Mix. The same samples were amplified using a kit from Vendor K according to the supplier’s instructions.
M: O’GeneRuler Express DNA Ladder, +: Control reaction with purified DNA, –: No-template control
Phusion Blood Direct PCR Master Mix is designed for amplification of DNA from whole blood at blood concentrations of up to 40% in the PCR reaction. This is achieved with specially modified Phusion Hot Start II High-Fidelity DNA Polymerase, which is tolerant to PCR inhibitors present in blood, as well as to most widely used blood preservatives (e.g., heparin, EDTA, citrate).
Figure 4. DNA was amplified from human and animal blood samples. Human blood preserved with (1) heparin, (2) citrate, (3) EDTA, or (4) stored on cards; (5) mouse; (6) cat; (7) dog; (8) cow; (9) bird; (10) sheep; (11) horse; and (12) fish.
M: O’GeneRuler Express DNA Ladder, +: Control reaction with purified DNA, –: No-template control
Phire Tissue Direct PCR Master Mix is designed for DNA amplification from animal and human tissues. The master mix contains Phire Hot Start II DNA Polymerase and is specially formulated for performing PCR in the presence of different tissue-derived inhibitors such as collagen, melanin and eumelanin (hair, skin), or myoglobin (muscle). The master mix is supplied with Thermo Scientific DNARelease Additive, which can be used to improve the release of DNA from difficult tissues.
Figure 2. DNA fragments from different human and animal tissue samples were amplified with Phire Tissue Direct PCR Master Mix. Mouse (1) tail, (2) ear, and (3) hair; bird (4) muscle and (5) feather; human (6) hair, (7) tooth, (8) nail, and (9) saliva; (10) zebrafish muscle; and (11) fruit fly.
M: Thermo Scientific O’GeneRuler Express DNA Ladder, +: Positive control reaction, –: No-template control
Phire Plant Direct PCR Master Mix is designed to amplify DNA directly from plant samples. The master mix contains Phire Hot Start II DNA Polymerase and is specially formulated for performing PCR from samples with plant-derived inhibitors such as complex polysaccharides and polyphenols.
Figure 3. Amplification of a 0.5 kb DNA fragment directly from leaves and seeds from different plants. A 0.5 mm punch of leaf (L) or a small piece of crushed seed (S) was placed directly into 50 μL of Phire Plant Direct PCR Master Mix. The same samples were amplified using a kit from Vendor K according to the supplier’s instructions.
M: O’GeneRuler Express DNA Ladder, +: Control reaction with purified DNA, –: No-template control
Phusion Blood Direct PCR Master Mix is designed for amplification of DNA from whole blood at blood concentrations of up to 40% in the PCR reaction. This is achieved with specially modified Phusion Hot Start II High-Fidelity DNA Polymerase, which is tolerant to PCR inhibitors present in blood, as well as to most widely used blood preservatives (e.g., heparin, EDTA, citrate).
Figure 4. DNA was amplified from human and animal blood samples. Human blood preserved with (1) heparin, (2) citrate, (3) EDTA, or (4) stored on cards; (5) mouse; (6) cat; (7) dog; (8) cow; (9) bird; (10) sheep; (11) horse; and (12) fish.
M: O’GeneRuler Express DNA Ladder, +: Control reaction with purified DNA, –: No-template control
Did you know about a universal direct PCR master mix that works with different sample types? It further simplifies and adds convenience to the direct PCR workflow.
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