Product Class: Other

Bst DNA Polymerase, Large Fragment
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

The original polymerase for Loop-Mediated Isothermal Amplification (LAMP)

  • Modified to retain 5´ → 3´ polymerase activity while lacking 5´ →3´ exonuclease activity
  • Suitable for applications requiring thermophilic strand displacement
  • Supplied with ThermoPol® Reaction Buffer for high product yields in demanding conditions
Catalog # Size Concentration
M0275S 1600.0 units 8000 units/ml
M0275L 8000.0 units 8000 units/ml
M0275M 8000.0 units 120000 units/ml

Product Information

Description

Highlights

  • Isolated from a recombinant source
  • Sequencing through problematic secondary structures
  • Supplied with 10X Reaction Buffer





Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the
5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity.

Product Source

Bst Polymerase, Large Fragment is prepared from an E. coli strain containing a genetic fusion of the Bacillus stearothermophilus DNA Polymerase gene, lacking the 5´ → 3´ exonuclease domain, and the gene coding for E. coli maltose binding protein (MBP). The fusion protein is purified to near homogeneity and the MBP portion of the fusion is cleaved off in vitro. The remaining polymerase is purified free of MBP (1).
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 at the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.

Reaction Conditions

1X ThermoPol® Reaction Buffer
Incubate at 65°C

1X ThermoPol® Reaction Buffer
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)

Activity in NEBuffers

NEBuffer™ 1: 50%
NEBuffer™ 2: 100%
NEBuffer™ 3: 50%
NEBuffer™ 4: 100%

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

Molecular Weight

Theoretical: 67000 daltons

5' - 3' Exonuclease

No

3' - 5' Exonuclease

No

Strand Displacement

++++

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)
  • DNA sequencing through high GC regions (2,3)
  • Rapid Sequencing from nanogram amounts of DNA template (4)

Product Notes

  1. Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
  2. 100 µg/ml BSA or 0.1%Triton X-100 is required for long term storage.
  3. Reaction temperatures above 70°C are not recommended.
  4. Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing or PCR.

References

  1. Kong, H., Aliotta, J. and Pelletier, J.J. New England Biolabs. Unpublished observation
  2. Griffin, H. and Griffin, A. (1994). PCRTechnology. 228-229.
  3. McClary, J. et al. (1991). J. DNASequencing and Mapping. 1, 173-180.
  4. Mead, D.A. et al. (1991). Biotechniques. 11, 76-87.
  5. Notomi et al. (2000). Nucleic Acids Res. 28(12), E63.
  6. Hsieh et al. (2014). Chem. Commun. 50, 3747-3749.
  7. Tanner and Evans (2014). Curr. Prot. Mol. Biol. 105, 15.14.

Protocols, Manuals & Usage

Protocols

  1. Typical LAMP Protocol (M0275)

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Can Bst DNA Polymerase be used in other NEBuffers, including rCutSmart?
  2. Can Bst DNA Polymerase be used to blunt DNA?
  3. Can Bst DNA Polymerase be used to fill in 3' overhangs?
  4. Can Bst DNA Polymerase be used to remove 5' overhangs?
  5. Can Bst DNA Polymerase be heat inactivated?
  6. Are NEB DNA Polymerases supplied with dNTPs?
  7. What are the main causes of reaction failure using Bst DNA Polymerase?
  8. Does Bst DNA Polymerase have an active 3'→5' proofreading exonuclease?
  9. Can Bst DNA Polymerase be used for thermal cycle sequencing?
  10. Can Bst DNA Polymerase initiate at a nick in the DNA?
  11. Can Bst DNA Polymerase be used in labeling reactions and partial fill in reactions?
  12. Can Bst DNA Polymerase be diluted?
  13. When should Bst DNA Polymerase be the enzyme of choice?
  14. Can Bst DNA Polymerase be used at temperatures other than 65°C?
  15. Does Bst DNA Polymerase, Large Fragment incorporate dUTP?
  16. Does Bst DNA polymerase have reverse transcriptase activity?
  17. Does NEB have a master mix for LAMP or RT-LAMP reactions?
  18. What is LAMP and RT-LAMP?
  19. What is the difference between Bst DNA Polymerase, Large Fragment and Bst 2.0® DNA Polymerase?

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

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.

Trademarks


NEW ENGLAND BIOLABS® and THERMOPOL® are registered trademarks of New England Biolabs, Inc.

TRITON® is a registered trademark of Union Carbide Corporation.

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