Direct PCR—Phire and Phusion Kits & Master Mixes

Mouse, plant, hair, and blood samples for direct PCR

Direct to PCR with fewer steps to discovery

The direct PCR approach delivers convenience for DNA amplification by allowing PCR directly from samples without prior DNA purification. Significant savings in time and cost can be achieved in applications such as plant and animal genotyping. We offer Thermo Scientific Phire and Phusion direct PCR products for fewer steps to your discovery. Explore our direct PCR master mixes and kits available based on sample types.

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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

The quality of PCR results was as good or better than with more time-consuming extraction methods, followed by PCR.

– Principal Investigator, Department of Biological Sciences, University of Manitoba, Winnipeg, Canada

By side stepping the need to extract and clean up the DNA we can save up to half a day using the Direct PCR kit, great when we are working with a number of different strains.

– Research Technician, Nematode Biology Research, Bristol, UK

Cleanest genotyping results I’ve ever had, in the shortest time, with the least amount of work.

– Research Assistant, Institute of Child Health, University College London, UK

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Two short protocols to suit your needs

Direct protocol and dilution and storage protocol of Thermo Scientific direct PCR kits

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.

Gel of direct PCR with human and animal 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

Animal and human tissues

Plant materials

Blood samples

Different sample types for Platinum direct PCR Universal Master Mix

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.

Learn more