DNA Amplification & PCR
Phusion® High-Fidelity
DNA Polymerase
DNA Cloning
Phusion Hot Start
Flex DNA Polymerase
RNA Analysis
DNA Amplification & PCR
Epigenetics
®
Sample Prep for Next Gen Sequencing
Protein Expression & Analysis
Phusion® High-Fidelity DNA Polymerase offers robust performance for a wide range
of templates. Its unique structure includes a novel Pyrococcus-like enzyme fused to a
processivity-enhancing domain; this combination enhances fidelity, speed and overall
performance. Reagent selection includes a standalone enzyme, master mix and kit
format, as well as a choice of reaction buffers for amplification of difficult templates.
Phusion Hot Start Flex DNA Polymerase is available as standalone enzyme or in a
master mix format, and enables high specificity amplification of a broad range of
templates. Phusion DNA Polymerase is a superior choice for cloning and sets the
standard for high-fidelity PCR polymerases.
Due to its enhanced processivity, Phusion DNA Polymerase generates PCR products with much
shorter reaction times. Extension times are reduced to 15-30 seconds per kilobase, while processivity is 10-fold greater than Pyrococcus furiosus (Pfu) DNA Polymerase and twice that of Taq DNA
Polymerase.
The fidelity of Phusion DNA Polymerase is 50X higher than Taq DNA Polymerase and
8.3X higher than Pyrococcus furiosus (Pfu) DNA Polymerase, making it the most accurate thermostable polymerase available.
Phusion DNA Polymerase generates robust amplification even with short
extension times.
• Robust Reactions – Maximal success
with minimal optimization
• Extreme Fidelity – Highest of any
available thermostable polymerase
(50X greater than Taq)
• High Speed – Dramatically reduced
extension times
High Fidelity Pfu
30s2m
• Versatile – Can be used for routine PCR
as well as long or difficult templates
• Hot Start – Flexible formulation allows
room temperature reaction setup and no
activation step
• Ease-of-Use – 1 unit per 50 µl reaction
Phusion DNA Polymerase has the highest
fidelity
50x
Modified KOD
30s2m
4,900
2,900
1,900
1,100
700
500
Fidelity (relative to Taq)
Phusion from NEB
30s2m
Advantages:
• High Yield – Increased product yield
using minimal amount of enzyme
Speed & Fidelity
bp
Cellular Analysis
6x
300
1x
Phusion
Pfu
Taq
Fidelity assays were performed using a lacI-based
method modified from Frey & Suppman, 1995.
100
A 1.2 kb C. elegans genomic amplicon was analyzed using different polymerases. Reactions were set up in duplicate and visualized
after microfluidic LabChip® analysis. All reactions were conducted according to manufacturer’s instructions using 30 cycles and
extension times of either 30 seconds or 2 minutes as indicated.
PCR_PHU_MS 11/11
DNA amplification & pcr
Phusion Hot Start Flex DNA Polymerase
Tips:
Phusion Hot Start Flex DNA Polymerase delivers additional flexibility to your reaction setup
and cycling conditions. In contrast to chemically modified or antibody-based hot start polymerases, Phusion Hot Start Flex utilizes an innovative synthetic aptamer. This structure binds
to the polymerase through non-covalent interactions, blocking activity during reaction setup.
The polymerase is activated during normal cycling conditions, allowing reactions to be set up at
room temperature. Phusion Hot Start Flex does not require a separate high temperature incubation step, shortening reaction times and increasing ease-of-use. Phusion Hot start Flex is an ideal
choice for high specificity amplification over a flexible range of annealing temperatures and
offers robust amplification on a wide variety of templates.
• 0.5–1.0 units of Phusion per 50 µl reaction
volume is recommended. Do not use more
than 2 units/50 µl reaction.
Phusion Hot Start Flex DNA Polymerase delivers robust amplification for a
wide range of templates.
1.3 kb
P
bp
4,900
2.3 kb
C
A
M
P
M
C
P
C
• To determine the optimal annealing
temperatures for a given set of primers,
use of the NEB Tm Calculator is
highly recommended (www.neb.com/
TmCalculator).
• An extension time of 15–30 sec/kb is
recommended. Do not use extension times
greater than 1 min/kb.
3.9 kb
A
• Typically, 0.5 µM of each primer is optimal
for Phusion. Phusion will not incorporate
dUTP.
A
M
• A denaturation temp of 98°C is
recommended.
2,900
1,900
P = Phusion Hot Start Flex
1,100
C = Chemical-based
Hot Start Polymerase
700
500
• Phusion DNA Polymerase will generate
blunt ended products.
A = Antibody-based
Hot Start Polymerase
1. Frey & Suppman (1995) Biochemica, 2, 34–35.
300
All amplicons are from human Jurkat template except for the 1.3 kb C. elegans amplicon. Amplicon sizes are indicated above gel. All reactions
were conducted in duplicate according to manufacturer’s instructions using 30 cycles and visualized after microfluidic LabChip® analysis.
Please note that Phusion product numbers
have changed (e.g. F-530 is now M0530).
Visit www.neb.com/phusion for more information.
Note that pack sizes will remain the same.
Ordering Information
Product
NEB #
SIZE
Phusion® High-Fidelity DNA Polymerase
M0530S/L
100/500 units
Phusion® Hot Start Flex DNA Polymerase
M0535S/L
100/500 units
Phusion Hot Start Flex 2X Master Mix
M0536S/L
100/500 rxns (50 µl vol)
Phusion® High-Fidelity PCR Master Mix with HF Buffer
M0531S/L
100/500 rxns (50 µl vol)
Phusion® High-Fidelity PCR Master Mix with GC Buffer
M0532S/L
100/500 rxns (50 µl vol)
Phusion® PCR Kit
E0553S/L
50/200 rxns (50 µl vol)
®
COMPANION PRODUCTS
Phusion® HF Buffer Pack
B0518S
6.0 ml
Phusion® GC Buffer Pack
B0519S
6.0 ml
Detergent-free Phusion® HF Buffer Pack
B0520S
6.0 ml
Detergent-free Phusion GC Buffer Pack
B0521S
6.0 ml
®
Notice to Purchaser: Limited License.
The purchase price of this product includes a limited, non-transferable
license under U.S. and foreign patents owned by BIO-RAD Laboratories,
Inc., to use this product. No other license under these patents is conveyed
expressly or by implication to the purchaser by the purchase of this
product.
The purchase price of this product includes a limited, non-transferable
license under a patent owned by New England Biolabs, Inc., to use this
product. No other license under this patent is conveyed expressly or by
implication to the purchaser by the purchase of this product.
Nucleic acid-based aptamers for use with thermophilic DNA polymerases
are licensed exclusively by New England Biolabs, Inc. from SomaLogic,
Inc. (See Patent Nos. 5,475,096; 5,670,637; 5,696,249; 5,874,557;
and 5,693,502). New England Biolabs, Inc. gives the Buyer/User a nonexclusive license to use the aptamer-based Phusion Hot Start Flex DNA
Polymerase for RESEARCH PURPOSES ONLY. Commercial use of the
aptamer-based Phusion Hot Start Flex DNA Polymerase requires a license
from New England Biolabs, Inc. Please contact [email protected] for more
information.
LabChip ® is a registered trademark of Caliper Life Sciences, Inc.
Phusion DNA Polymerase was developed by Finnzymes Oy, now a part of Thermo Fisher Scientific. This product is manufactured by
New England Biolabs, Inc. under agreement with, and under the performance specifications of Thermo Fisher Scientific.
Phusion® is a registered trademark and property of Thermo Fisher Scientific.
www.neb.com
New England Biolabs (UK) Ltd.
www.neb.uk.com
New England Biolabs France
New England Biolabs GmbH, Germany
www.neb-online.fr
www.neb-online.de
For a complete list of international offices and distributors, please visit www.neb.com