Phusion^® Hot Start Flex 2X Master Mix

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0536S     -20       Phusion® Hot Start Flex 2X Master Mix M0536SVIAL -20 2 x 1.25 ml 2 X   DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
M0536L     -20       Phusion® Hot Start Flex 2X Master Mix M0536SVIAL -20 10 x 1.25 ml 2 X   DMSO B0515AVIAL -20 2 x 0.5 ml 100 %

Application Features

• High-specificity PCR
• Cloning
• Long or Difficult Amplification
• High-Throughput PCR

Companion Products

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

1. Phusion Hot Start Flex DNA Polymerase is unlike other enzymes and care must be taken when designing cycling protocols. To determine the optimal annealing temperatures for a given set of primers, use of the NEB T_m Calculator is highly recommended.
2. Product specifications for individual components in the Phusion Hot Start Flex 2X Master Mix are available separately.

1. Frey, B. and Suppman, B. (1995). BioChemica. 2, 34-35.
2. Chester, N. and Marshak, D.R. (1993). Analytical Biochemistry. 209, 284-290.

1. Protocol for Phusion® Hot Start Flex 2X Master Mix

• Activity of Restriction Enzymes in PCR Buffers

• DNA Polymerase Selection Chart
• Thermophilic DNA Polymerases

• PCR Selector
• Tm Calculator

1. What are the differences between the various available Phusion^® products?
2. My 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. What are Hot Start and WarmStart^® polymerases and when would I use them?

• PCR Troubleshooting Guide

• Polymerase Fidelity: What is it, and what does it mean for your PCR?
• Using aptamers to control enzyme activities: Hot Start Taq and beyond

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]

• M0536S_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]

• M0536S_L_v1_0211610
• M0536S_L_v2_0251712
• M0536L_v2_10009797
• M0536S_v2_10017191
• M0536L_v2_10022234
• M0536S_v2_10021901
• M0536L_v2_10034378
• M0536L_v2_10036746
• M0536S_v2_10040453
• M0536L_v2_10043252
• M0536L_v2_10045706
• M0536L_v2_10047106
• M0536S_v2_10048081
• M0536S_v2_10050356
• M0536S_v2_10053908
• M0536L_v2_10050359
• M0536S_v2_10057676
• M0536L_v2_10057681
• M0536S_v2_10061426
• M0536L_v2_10063540
• M0536S_v2_10064552
• M0536L_v2_10065362
• M0536S_v2_10065365
• M0536L_v2_10073014
• M0536L_v2_10077359
• M0536S_v2_10079528
• M0536L_v2_10079527
• M0536S_v2_10084952
• M0536L_v2_10085281
• M0536L_v2_10088164
• M0536S_v2_10091661
• M0536S_v2_10095576
• M0536L_v2_10095575
• M0536L_v2_10107308
• M0536S_v2_10107311
• M0536L_v2_10111149
• M0536S_v2_10110670
• M0536L_v2_10117356
• M0536S_v2_10114453
• M0536S_v2_10122343
• M0536L_v2_10122827
• M0536L_v2_10128114
• M0536S_v2_10126949
• M0536S_v2_10134159
• M0536L_v2_10132982
• M0536S_v2_10140980
• M0536L_v2_10144284
• M0536S_v2_10144283
• M0536L_v2_10150890
• M0536S_v2_10151866
• M0536L_v2_10155852
• M0536S_v2_10156022
• M0536S_v2_10163723
• M0536L_v2_10162791
• M0536S_v2_10166267
• M0536L_v2_10171895
• M0536L_v2_10177161
• M0536S_v2_10174497
• M0536S_v2_10184154
• M0536L_v2_10184155
• M0536L_v2_10191815
• M0536S_v2_10187398
• M0536S_v2_10196303
• M0536S_v2_10205309
• M0536L_v2_10206441
• M0536L_v2_10220969
• M0536S_v2_10220968
• M0536L_v2_10226673
• M0536S_v2_10226672
• M0536L_v2_10233795
• M0536S_v2_10233794

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

• Phusion^® Hot Start Flex 2X Master Mix
• DMSO

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

Notice to Customers: Phusion^® DNA Polymerase was developed by Finnzymes Oy, now a part of Thermo Fisher Scientific.

Phusion^® is a registered trademark and property of Thermo Fisher Scientific.

Notice to Purchaser: Limited License. The purchase price of this product includes a limited, non-transferable license under U.S. and foreign patent 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.

Trademarks

NEW ENGLAND BIOLABS^® and Q5^® are registered trademarks of New England Biolabs, Inc.
PHUSION^® is a registered trademark and property of Thermo Fisher Scientific.
THERMO SCIENTIFIC^® is a registered trademark of Thermo Fisher Scientific.