Phusion^™ High-Fidelity PCR Kit

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
NEB # Stored at (°C) E0553S -20 Component Name Component # Stored at (°C) Amount Concentration Phusion™ High-Fidelity DNA Polymerase M0530AVIAL -20 1 x 0.025 ml 2,000 units/ml Phusion™ HF Buffer Pack B0518SVIAL -20 1 x 1.5 ml 5 X Phusion™ GC Buffer Pack B0519SVIAL -20 1 x 1.5 ml 5 X Deoxynucleotide (dNTP) Solution Mix N0447AVIAL -20 1 x 0.1 ml 10 mM each dNTP MgCl2 solution B0510AVIAL -20 1 x 1.5 ml 50 mM Control Lambda Template N3010AVIAL -20 1 x 0.04 ml 0.5 ng/µl 1.3 kb Control Primer Mix S0535AVIAL -20 1 x 0.05 ml 4 µM 10 kb Control Primer Mix S0536AVIAL -20 1 x 0.05 ml 4 µM DMSO B0515AVIAL -20 1 x 0.5 ml 100 % Quick-Load DNA Marker, Broad Range N0303AVIAL -20 1 x 0.2 ml 100 µg/ml
NEB # Stored at (°C) E0553L -20 Component Name Component # Stored at (°C) Amount Concentration Phusion™ High-Fidelity DNA Polymerase M0530AAVIAL -20 1 x 0.1 ml 2,000 units/ml Phusion™ HF Buffer Pack B0518SVIAL -20 3 x 1.5 ml 5 X Phusion™ GC Buffer Pack B0519SVIAL -20 1 x 1.5 ml 5 X Deoxynucleotide (dNTP) Solution Mix N0447AAVIAL -20 1 x 0.4 ml 10 mM each dNTP MgCl2 solution B0510AVIAL -20 1 x 1.5 ml 50 mM Control Lambda Template N3010AVIAL -20 1 x 0.04 ml 0.5 ng/µl 1.3 kb Control Primer Mix S0535AVIAL -20 1 x 0.05 ml 4 µM 10 kb Control Primer Mix S0536AVIAL -20 1 x 0.05 ml 4 µM DMSO B0515AVIAL -20 1 x 0.5 ml 100 % Quick-Load DNA Marker, Broad Range N0303AAVIAL -20 1 x 0.4 ml 100 µg/ml

Features

Application features:

• PCR
• Primer Extension
• Cloning
• Longer Difficult Amplification
• High-Throughput PCR

Companion Products

• Deoxynucleotide (dNTP) Solution Set
• Magnesium Chloride (MgCl₂) Solution
• Phusion™ High-Fidelity PCR Master Mix with HF Buffer
• Phusion™ High-Fidelity PCR Master Mix with GC Buffer
• Phusion™ Hot Start Flex DNA Polymerase
• Phusion™ Hot Start Flex 2X Master Mix

Materials Sold Separately

• Phusion™ High-Fidelity DNA Polymerase
• Deoxynucleotide (dNTP) Solution Mix
• Lambda DNA

1. Saiki, R.K. et al. (1985). Science. 230, 1350-1354.
2. Chien, A., Edgar, D.B. and Trela, J.M. (1976). J. Bact.. 127, 1550-1557.
3. Kaledin, A.S., Slyusarenko, A.G. and Gorodetskii, S.I. (1980). Biokhimiya. 45, 644-651.
4. Lawyer, F.C. et al. (1993). PCR Method and Appl.. 2, 275-287.
5. Longley, M.J., Bennett, S.E. and Mosbaugh D.W. (1990). Nucleic Acids Res.. 18, 7317-7322.
6. Lyamichev, V., Brow, M.A. and Dahlberg, J.E. (1993). Science. 260, 778-783.
7. Frey and Suppman (1995). Biochimica. 34-35.
8. Breslauer, et al. (1986). PNAS. 83, 3746-3750.

1. PCR Optimization of the Control Template using Phusion™ High-Fidelity PCR Kit
2. Protocol for a Routine PCR with Phusion™ High-Fidelity PCR Kit
3. PCR Optimization with Phusion™ High-Fidelity PCR Kit

• Activity of Restriction Enzymes in PCR Buffers
• Guidelines for PCR Optimization with Thermophilic DNA Polymerases

• DNA Polymerase Selection Chart
• Thermophilic DNA Polymerases

• Tm Calculator

1. What are the differences between the various available Phusion™ products?
2. My PCR results are not as expected. Where can I find troubleshooting help?
3. What ends will my PCR products have?
4. What does exonuclease activity mean for a DNA polymerase?
5. Does Phusion™ High-Fidelity DNA Polymerase exhibit a strand displacement activity?
6. What are the properties of this polymerase (fidelity, product ends, max amplicon, modified base incorporation, etc.)?
7. How should I determine the appropriate annealing temperature for my reaction?
8. What should the final primer concentration be in my PCR?
9. My template is GC-rich or supercoiled. How can I optimize my product yield using Phusion™ High-Fidelity DNA Polymerase?
10. What are the details of the Quick-Load, Broad Range DNA Marker (NEB #N0303)?

• PCR Troubleshooting Guide

• Polymerase Fidelity: What is it, and what does it mean for your PCR?

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• E0553S_L_v2

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• E0553S_L_v1_0151708
• E0553S_v1_10013495
• E0553L_v1_10013926
• E0553S_v1_10021738
• E0553L_v1_10021732
• E0553S_v1_10023549
• E0553L_v1_10034492
• E0553S_v1_10034491
• E0553L_v1_10041340
• E0553S_v1_10041432
• E0553L_v1_10045328
• E0553S_v1_10047984
• E0553L_v1_10048545
• E0553L_v1_10050935
• E0553S_v1_10054175
• E0553S_v1_10055991
• E0553L_v1_10055985
• E0553L_v1_10060090
• E0553S_v1_10061490
• E0553L_v1_10063721
• E0553S_v1_10064331
• E0553S_v1_10065626
• E0553L_v1_10069926
• E0553S_v1_10073566
• E0553L_v1_10077741
• E0553S_v1_10079225
• E0553L_v1_10082375
• E0553L_v1_10084849
• E0553L_v1_10086366
• E0553S_v1_10086370
• E0553L_v1_10090571
• E0553S_v1_10091455
• E0553L_v1_10095925
• E0553S_v1_10095929
• E0553L_v1_10099933
• E0553S_v1_10099932
• E0553L_v1_10104358
• E0553S_v1_10104881
• E0553L_v1_10107125
• E0553S_v1_10108362
• E0553L_v1_10109593
• E0553S_v1_10113742
• E0553L_v1_10117360
• E0553S_v1_10115049
• E0553S_v1_10120996
• E0553L_v1_10120995
• E0553L_v1_10123923
• E0553L_v1_10125446
• E0553L_v1_10131931
• E0553L_v1_10132254
• E0553S_v1_10127745
• E0553S_v1_10137416
• E0553L_v1_10140019
• E0553L_v1_10145819
• E0553S_v1_10145487
• E0553L_v1_10150541
• E0553S_v1_10150731
• E0553L_v1_10153111
• E0553S_v1_10157016
• E0553L_v1_10157024
• E0553S_v1_10161330
• E0553L_v1_10161329
• E0553L_v1_10164750
• E0553S_v1_10166184
• E0553L_v1_10166359
• E0553S_v1_10170332
• E0553L_v1_10173613
• E0553S_v1_10176244
• E0553L_v1_10178503
• E0553L_v1_10182545
• E0553S_v1_10183504
• E0553L_v1_10192581
• E0553S_v1_10192582
• E0553L_v1_10203990
• E0553S_v1_10203989
• E0553S_v1_10208172
• E0553S_v1_10211321
• E0553S_v1_10216377
• E0553L_v1_10220972
• E0553S_v1_10221503
• E0553L_v1_10227768
• E0553S_v1_10236032
• E0553L_v1_10235649
• E0553S_v1_10238409
• E0553L_v1_10242787
• E0553S_v1_10245731
• E0553L_v1_10252442
• E0553S_v1_10252443
• E0553L_v1_10257556
• E0553S_v1_10264455
• E0553L_v1_10270571
• E0553S_v1_10276420
• E0553L_v1_10278053
• E0553S_v1_10280631
• E0553S_v2_10287176
• E0553L_v2_10281688
• E0553S_v2_10297866
• E0553L_v2_10298566
• E0553S_v2_10300302
• E0553L_v2_10308397
• E0553L_v2_10310158
• E0553L_v2_10319015
• E0553S_v2_10316624
• E0553S_v2_10327979
• E0553L_v2_10329598
• E0553S_v2_10338592
• E0553L_v2_10341853

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• Phusion^™ High-Fidelity DNA Polymerase
• Phusion^™ HF Buffer Pack
• Phusion^™ GC Buffer Pack
• Deoxynucleotide (dNTP) Solution Mix
• MgCl₂ solution
• Control Lambda Template
• 1.3 kb Control Primer Mix
• 10 kb Control Primer Mix
• DMSO
• Quick-Load DNA Marker, Broad Range

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

Licenses

Phusion™ DNA Polymerase was developed by Finnzymes Oy, now a part of Thermo Fisher Scientific. 
PHUSION™ and THERMO SCIENTIFIC™ are trademarks and property of Thermo Fisher Scientific.

Notice to purchaser: 
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.