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Bst 3.0 DNA Polymerase

Catalog # Concentration Size List Price Quantity Your Price
M0374L 8000 units/ml 8000 units $461.00
$414.90
M0374M 120000 units/ml 8000 units $461.00
$414.90
M0374S 8000 units/ml 1600 units $116.00
$104.40
Catalog # Size List Price Your Price
M0374L 8000 units $461.00
$414.90
M0374M 8000 units $461.00
$414.90
M0374S 1600 units $116.00
$104.40
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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
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    Lyophilizing reagents: requirements, challenges, and considerations for assay developers



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.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Bst 3.0 DNA Polymerase M0374SVIAL -20 1 x 0.2 ml 8,000 units/ml
  Isothermal Amplification Buffer II Pack B0374SVIAL -20 1 x 1.5 ml 10 X
  Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
  Bst 3.0 DNA Polymerase M0374LVIAL -20 1 x 1 ml 8,000 units/ml
  Isothermal Amplification Buffer II Pack B0374SVIAL -20 2 x 1.5 ml 10 X
  Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
  Bst 3.0 DNA Polymerase M0374MVIAL -20 1 x 0.067 ml 120,000 units/ml
  Isothermal Amplification Buffer II Pack B0374SVIAL -20 2 x 1.5 ml 10 X
  Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
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

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.

Companion Products
Notes
  • Bst 3.0 DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
  • Reaction temperatures above 72°C are not recommended.
  • Bst 3.0 DNA Polymerase cannot be used for PCR or reactions requiring thermal denaturation.
Citations
Additional Citations
  • Lee D, Shin Y, Chung S, Hwang KS, Yoon DS, Lee JH (2016) Simple and Highly Sensitive Molecular Diagnosis of Zika Virus by Lateral Flow Assays. Anal ChemPubMedID: 28193014, DOI: 10.1021/acs.analchem.6b03460
Quality Control Assay
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 Data Sheets
Legal And Disclaimer

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.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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