Product Class: Other

Bst 2.0 WarmStart® DNA Polymerase
NEBU cloned at NEB recombinant 65 80 Heat hotstart

Need assistance designing LAMP primers? Use the NEB LAMP Primer Design Tool.

This product is available in a glycerol-free format.

Product Introduction

Optimized for Loop-Mediated Isothermal Amplification (LAMP)

  • Consistent amplification performance with the convenience of room temperature setup
  • WarmStart® technology eliminates off-target amplification and offers increased reaction efficiency
  • Flexible reaction conditions including a higher salt tolerance and thermostability, as compared to wild-type Bst DNA Polymerase, Large Fragment
  • Optimal reaction performance from 60-72 °C
  • One of several products available glycerol-free
Catalog # Size Concentration
M0538S 1600.0 units 8000 units/ml
M0538L 8000.0 units 8000 units/ml
M0538M 8000.0 units 120000 units/ml

Product Information

Description




Bst 2.0 WarmStart DNA Polymerase is an in silico designed homologue of Bacillus stearothermophilus DNA Polymerase I, Large Fragment (Bst DNA Polymerase, Large Fragment) with a reversibly-bound aptamer, which inhibits polymerase activity at temperatures below 45°C. The aptamer rapidly releases the Bst 2.0 WarmStart DNA Polymerase above 45°C and therefore no special activation step is needed to activate the polymerase. Bst 2.0 WarmStart DNA Polymerase contains 5´→3´ DNA polymerase activity and strong strand-displacement activity but lacks 5´→3´ exonuclease activity. Bst 2.0 WarmStart DNA Polymerase displays improved amplification speed, yield, salt tolerance, and thermostability compared to wild-type Bst DNA Polymerase, Large Fragment.

To support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows, Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) is also offered in a glycerol-free format.

Benefits of Bst 2.0 WarmStart Benefits of Bst 2.0 WarmStart

Identical LAMP reactions were run either immediately after setup (solid line) or after a 2 hour incubation at 25 °C. Without the protection from Bst 2.0 WarmStart, this room temperature incubation results in variable LAMP performance. Bst 2.0 WarmStart provides more consistent amplification reaction and enables room-temperature and high-throughput setup.

Product Source

Bst 2.0 WarmStart DNA Polymerase is prepared from an E. coli strain that expresses the Bst 2.0 DNA Polymerase protein from an inducible promoter.
This product is related to the following categories:
Isothermal Amplification & Strand Displacement Products,
This product can be used in the following applications:
Whole Genome Amplification,
Strand Displacement Amplification & Nicking Enzyme,
Loop-Mediated Isothermal Amplification,
Isothermal Amplification

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 25 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.

Reaction Conditions

1X Isothermal Amplification Buffer Pack
Incubate at 65°C

1X Isothermal Amplification Buffer Pack
20 mM Tris-HCl
10 mM (NH4)2SO4
50 mM KCl
2 mM MgSO4
0.1% Tween® 20
(pH 8.8 @ 25°C)

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% Triton® X-100
pH 7.1 @ 25°C

Heat Inactivation

80°C for 20 minutes

Unit Assay Conditions

50 mM KCl, 20 mM Tris- HCl (pH 8.8), 10 mM MgCl2, 30 nM M13mp18 SS DNA, 70 nM M13 sequencing primer (–47) 24 mer, 200 μM dATP, 200 μM dCTP, 200 μM dGTP, 100 μM dTTP including [3H]-dTTP and 100 μg/ml BSA.

Application Features

  • Isothermal amplification (LAMP)
  • Applications requiring strand-displacement DNA synthesis
  • DNA sequencing through high GC regions
  • Rapid sequencing from nanogram amounts of DNA template

Product Notes

  1. Bst 2.0 WarmStart DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
  2. Reaction temperatures above 70°C are not recommended.
  3. Bst 2.0 WarmStart DNA Polymerase cannot be used for thermal cycle sequencing or PCR.
  4. Specific reaction conditions will vary for differentisothermal amplification applications. For best results, use 1X Isothermal Amplification Buffer.

Protocols, Manuals & Usage

Protocols

  1. Typical LAMP Protocol (M0538)
  2. Strand Displacement Amplification (SDA) Protocol using WarmStart® Nt.BstNBI (NEB #R0725)

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the difference between Bst DNA Polymerase, Large Fragment and Bst 2.0® DNA Polymerase?
  2. Why would I use Bst 2.0 WarmStart® DNA Polymerase?
  3. Can Bst DNA 2.0 Polymerase be used in other NEBuffers?
  4. Can Bst 2.0® DNA Polymerase be used to blunt DNA?
  5. Can Bst 2.0® DNA Polymerase be used to fill in 3' overhangs?
  6. Can Bst 2.0® DNA Polymerase be used to remove 5' overhangs?
  7. Can Bst 2.0® DNA Polymerase be heat inactivated?
  8. Are NEB DNA Polymerases supplied with dNTPs?
  9. What are the main causes of reaction failure using Bst 2.0® DNA Polymerase?
  10. Does Bst 2.0® DNA Polymerase have an active 3'→5' proofreading exonuclease?
  11. Can Bst 2.0® DNA Polymerase be used in applications requiring thermal cycling?
  12. Can Bst 2.0® DNA Polymerase initiate at a nick in the DNA?
  13. Can Bst 2.0® DNA Polymerase be used in labeling reactions and partial fill in reactions?
  14. Can Bst 2.0® DNA Polymerase be diluted?
  15. When should Bst 2.0® DNA Polymerase be the enzyme of choice?
  16. Can Bst 2.0® DNA Polymerase be used at temperatures other than 65°C?
  17. Does Bst 2.0® DNA polymerase incorporate dUTP?
  18. How do I use Antarctic Thermolabile UDG for carryover prevention in LAMP reactions?
  19. Does NEB have a master mix for LAMP or RT-LAMP reactions?
  20. What is LAMP and RT-LAMP?
  21. What are Hot Start and WarmStart® polymerases and when would I use them?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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.

Specifications

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]

Certificate Of Analysis

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]

Safety DataSheets

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

Legal and Disclaimers

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

Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the Bst 2.0 WarmStart DNA Polymerase for Research Use Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact busdev@neb.com. 

The purchase of NEB Bst products conveys to the purchaser the limited, nontransferable right to use the purchased products to perform loop-mediated isothermal amplification ("LAMP") for research use only.  LAMP is a patented technology belonging to Eiken Chemical Co., Ltd. and any use other than research may require a license from Eiken Chemical Co., Ltd.

Trademarks

NEW ENGLAND BIOLABS® and THERMOPOL® are registered trademarks of New England Biolabs, Inc.
WARMSTART® is a trademark of New England Biolabs, Inc.
TRITON® is a registered trademark of Union Carbide Corporation.
TWEEN® is a registered trademark of Uniqema Americas LLC.

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