Product Class: Other

Bst 3.0 DNA Polymerase
NEBU cloned at NEB recombinant 65 80 Heat

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

This product is available in a glycerol-free format. Contact us for more information.

Product Introduction

Bst 3.0 DNA Polymerase demonstrates robust performance even in high concentrations of amplification inhibitors and features significantly increased reverse transcriptase activity compared to Bst DNA Polymerase.

  • Fused to novel nucleic acid binding domain for enhanced performance
  • Optimized for Loop-Mediated Isothermal DNA Amplification (LAMP)
  • Simplified reaction setup: single-enzyme RT-LAMP reaction
  • High reverse transcriptase activity up to 72°C
  • Offers fast polymerization rates and robust performance even in presence of inhibitors, including dUTP
  • Learn more about LAMP and other isothermal amplification methods
Catalog # Size Concentration
M0374S 1600.0 units 8000 units/ml
M0374L 8000.0 units 8000 units/ml
M0374M 8000.0 units 120000 units/ml

Product Information

Description



Bst 3.0 DNA Polymerase is an in silico designed homologue of Bacillus stearothermophilus DNA Polymerase I, Large Fragment engineered and fused to a novel nucleic acid binding domain for improved isothermal amplification performance and increased reverse transcription activity. Bst 3.0 DNA Polymerase contains 5´→3´ DNA polymerase activity with either DNA or RNA templates and strong strand displacement activity, but lacks 5´→3´ and 3´→5´ exonuclease activity. Bst 3.0 DNA Polymerase demonstrates robust performance even in high concentrations of amplification inhibitors, including dUTP and features significantly increased reverse transcriptase activity compared to Bst DNA Polymerase.

An example of the utility and speed offered by Bst 3.0 DNA Polymerase can be seen in this publication that explores the development of a novel digital LAMP assay: Rapid pathogen-specific phenotypic antibiotic susceptibility testing using digital LAMP quantification in clinical samples.

Fast, single-enzyme RT-LAMP can be performed using Bst 3.0 
RT-LAMP was performed using indicated DNA polymerase and Jurkat total RNA and primers for two genes (ACTB, left; HMBS2, right). Fastest results were observed with a 2-enzyme system, Bst 2.0® and WarmStart RTx, but robust amplification was also observed using Bst 3.0 without additional RT. Bst LF, Bst 2.0 and competitor enzymes showed highly variable performance, with slow threshold times or reaction failure on one of the two targets.

Product Source

An E. coli strain that carries the engineered Bst 3.0 gene.
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,
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 II Pack
Incubate at 65°C

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

Activity in NEBuffers

rCutSmart™ Buffer: 100%
NEBuffer™ r1.1: 10%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 100%

Storage Buffer

100 mM KCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
0.1% Triton® X-100
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

80°C for 5 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, RT-LAMP)
  • Applications requiring strand-displacement DNA synthesis
  • Reverse transcription at elevated temperature (to 72°C)
  • Amplification in the presence of impure samples or amplification inhibitors

Product Notes

  1. Bst 3.0 DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
  2. Reaction temperatures above 72°C are not recommended.
  3. Bst 3.0 DNA Polymerase cannot be used for PCR or reactions requiring thermal denaturation.

Protocols, Manuals & Usage

Protocols

  1. Typical LAMP Protocol (M0374)

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the difference between Bst DNA Polymerase, Large Fragment, Bst 2.0®, and Bst 3.0 DNA Polymerase?
  2. Why would I use Bst 3.0 DNA Polymerase?
  3. Can Bst 3.0 DNA Polymerase be used in other NEBuffers?
  4. How active is Bst 3.0 at other temperatures?
  5. Does Bst 3.0 DNA polymerase incorporate dUTP?
  6. Does Bst 3.0 DNA polymerase have reverse transcriptase activity?
  7. Can Bst 3.0 DNA Polymerase be used to blunt DNA?
  8. Can Bst 3.0 DNA Polymerase be used to fill in 3' overhangs?
  9. Can Bst 3.0 DNA Polymerase be used to remove 5' overhangs?
  10. Can Bst 3.0 DNA Polymerase be heat inactivated?
  11. Are NEB DNA Polymerases supplied with dNTPs?
  12. What are the main causes of reaction failure using Bst 3.0 DNA Polymerase?
  13. Does Bst 3.0 DNA Polymerase have an active 3'→5' proofreading exonuclease?
  14. Can Bst 3.0 DNA Polymerase be used in applications requiring thermal cycling?
  15. Can Bst 3.0 DNA Polymerase initiate at a nick in the DNA?
  16. Can Bst 3.0 DNA Polymerase be used in labeling reactions and partial fill in reactions?
  17. Can Bst 3.0 DNA Polymerase be diluted?
  18. When I thaw Isothermal Amplification Buffer II I see a lot of white precipitate, is this normal?
  19. When should Bst 3.0 DNA Polymerase be the enzyme of choice?
  20. How do I reduce non-template amplification (NTC) in LAMP reactions with Bst 3.0?
  21. How do I use Antarctic Thermolabile UDG for carryover prevention in LAMP reactions?
  22. Does NEB have a master mix for LAMP or RT-LAMP reactions?
  23. What is LAMP and RT-LAMP?

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

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. A patent is pending for NEB's Bst DNA Polymerase.

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