Higher success and simpler setup with a trusted PCR enzymeThermo Scientific™ Phusion™ Plus DNA Polymerase is the latest addition to our family of Phusion™ products, which are Pyrococcus polymerase–like high-fidelity enzymes fused to a DNA-binding domain. This unique protein fusion technology enables Phusion Plus DNA Polymerase to generate PCR sequences with high accuracy, sensitivity, and inhibitor tolerance. In addition, its specialized reaction buffer allows PCR amplification without the need to calculate primer annealing temperatures, helping you save time and avoid mistakes in PCR runs.

Highlights• High fidelity—provides >100x higher sequence accuracy than Taq polymerase

• Convenient setup—allows for a universal annealing temperature of 60°C

• Enhanced specificity—reduces nonspecific amplification and primer degradation via hot-start modification

• Benchtop stability—enables lab automation setup since assembled reactions are stable at room temperature for 24 hours

• High inhibitor tolerance—works with DNA of suboptimal purity

For Research Use Only. Not for use in diagnostic procedures. © 2021 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Q5 is a trademark of New England Biolabs, Inc. COL113948 0221

Get more details at thermofisher.com/phusionplus

150

125

100

75

50

25

0

Phusion Plus e

nzyme

Q5 Hot Start H

igh-Fidelity enzym

e

Phusion Hot S

tart II enzym

e

PrimeSTA

R GXL enzyme

KAPA HiFi Hot S

tart enzym

e

Taq enzym

e

Figure 1. Fidelity relative to Taq DNA polymerase. Fidelity values were determined by next-generation sequencing with molecular barcodes. Phusion Plus DNA Polymerase generates PCR amplicons with very few errors, due to its extremely high fidelity >100x that of Taq enzyme.

Figure 3. Highly sensitive and specific PCR. A 0.5 kb target can be detected from as little as 0.08 ng of human gDNA using Phusion Plus DNA Polymerase.

Figure 5. Enzyme tolerance to common PCR inhibitors. A 2 kb target was amplified from human gDNA, with the reactions containing: (1) no inhibitor; (2) humic acid (0.5 µg/mL); (3) hemin (2.5 µM); (4) xylan (250 µg/mL). Phusion Plus DNA Polymerase (PP) amplified the targets more efficiently than Phusion™ Hot Start Flex DNA Polymerase (PF) and Q5 Hot Start High-Fidelity DNA Polymerase (Q5) in the presence of the inhibitors.

Figure 4. Reaction stability on the benchtop. Assembled PCR reactions were loaded immediately (0 hr) onto a thermal cycler or incubated at room temperature for 24 hours before thermal cycling. With a stringent hot start, of Phusion Plus DNA Polymerase (PP) produced PCR amplicons with high specificity and yields even after 24 hours, in contrast to NEB Q5™ Hot Start High-Fidelity DNA Polymerase (Q5).

Figure 2. Specific and efficient amplification using a universal annealing temperature. Targets of different lengths were amplified efficiently from human genomic DNA (gDNA) using Phusion Plus DNA Polymerase and an annealing temperature of 60°C. Calculated annealing temperatures of the primers range from 61°C to 69°C.

0.3

kb0.

7 kb

2.0

kb3.

9 kb

7.5 k

b 250ng

50ng

10ng

2.0ng

0.4ng

0.08ng

0ng

PP

0 hr 24 hr

Q5

0 hr 24 hr PP

1 2 3 4

Q5

1 2 3 4

PF

1 2 3 4

Ordering information

Description Size* Cat. No.

Phusion Plus DNA Polymerase 100 reactions F630S

Phusion Plus PCR Master Mix 100 reactions F631S

Phusion Plus Green PCR Master Mix 100 reactions F632S

* Additional product sizes available.

Visit our molecular biology resource library for scientific webinars, videos, articles, and more at thermofisher.com/mbresources

Using one cycling protocol

Fifth target (e.g., 7 kb)

Third target (e.g., 2 kb)

Second target (e.g., 1 kb)

Fourth target (e.g., 4 kb)

First target (e.g., 0.5 kb)

Save time by co-cycling PCR

targets of different lengths using

Phusion Plus DNA Polymerase

and its universal protocol.