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Bst DNA Polymerase, Large Fragment

Catalog # Concentration Size List Price Quantity Your Price
M0275L 8000 units/ml 8000 units $468.00
$421.20
M0275M 120000 units/ml 8000 units $468.00
$421.20
M0275S 8000 units/ml 1600 units $117.00
$105.30
Catalog # Size List Price Your Price
M0275L 8000 units $468.00
$421.20
M0275M 8000 units $468.00
$421.20
M0275S 1600 units $117.00
$105.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Bst DNA Polymerase, Large Fragment M0275SVIAL -20 1 x 0.2 ml 8,000 units/ml
  ThermoPol® Reaction Buffer B9004SVIAL -20 1 x 1.5 ml 10 X
  Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
  Bst DNA Polymerase, Large Fragment M0275LVIAL -20 1 x 1 ml 8,000 units/ml
  ThermoPol® Reaction Buffer B9004SVIAL -20 2 x 1.5 ml 10 X
  Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
  Bst DNA Polymerase, Large Fragment M0275MVIAL -20 1 x 0.067 ml 120,000 units/ml
  ThermoPol® Reaction Buffer B9004SVIAL -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)
  • DNA sequencing through high GC regions (2,3)
  • Rapid Sequencing from nanogram amounts of DNA template (4)

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

NEBufferâ„¢ 1: 50%
NEBufferâ„¢ 2: 100%
NEBufferâ„¢ 3: 50%
NEBufferâ„¢ 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.

Companion Products
Materials Sold Separately
Notes
  • Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
  • 100 µg/ml BSA or 0.1%Triton X-100 is required for long term storage.
  • Reaction temperatures above 70°C are not recommended.
  • Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing or PCR.
References
  • Kong, H., Aliotta, J. and Pelletier, J.J. New England Biolabs. Unpublished observation
  • Griffin, H. and Griffin, A. (1994). PCRTechnology. 228-229.
  • McClary, J. et al. (1991). J. DNASequencing and Mapping. 1, 173-180.
  • Mead, D.A. et al. (1991). Biotechniques. 11, 76-87.
  • Notomi et al. (2000). Nucleic Acids Res. 28(12), E63.
  • Hsieh et al. (2014). Chem. Commun. 50, 3747-3749.
  • Tanner and Evans (2014). Curr. Prot. Mol. Biol. 105, 15.14.
Citations
Additional Citations
  • Trisadee Khamlor, Petai Pongpiachan, Rangsun Parnpai, Kanchana Punyawai, Siwat Sangsritavong, Nipa Chokesajjawatee (2015) Bovine embryo sex determination by multiplex loop-mediated isothermal amplification. Theriogenology 83, 891-6.PubMedID: 25542460, DOI: 10.1016/j.theriogenology.2014.11.025
  • DoKyung Lee, Eun Jin Kim, Paul E Kilgore, Soon Ae Kim, Hideyuki Takahashi, Makoto Ohnishi, Dang Duc Anh, Bai Qing Dong, Jung Soo Kim, Jun Tomono, Shigehiko Miyamoto, Tsugunori Notomi, Dong Wook Kim, Mitsuko Seki (2015) Clinical Evaluation of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Neisseria meningitidis in Cerebrospinal Fluid. PLoS One 10, e0122922.PubMedID: 25853422, DOI: 10.1371/journal.pone.0122922
  • Manabu Nemoto, Yoshinori Morita, Hidekazu Niwa, Hiroshi Bannai, Koji Tsujimura, Takashi Yamanaka, Takashi Kondo (2015) Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods 215-216, 13-6.PubMedID: 25682750, DOI: 10.1016/j.jviromet.2015.02.001
  • Mohammad Reza Allahyar Torkaman, Kazunari Kamachi, Vajihe Sadat Nikbin, Masoumeh Nakhost Lotfi, Fereshteh Shahcheraghi (2015) Comparison of loop-mediated isothermal amplification and real-time PCR for detecting Bordetella pertussis. J Med Microbiol 64, 463-5.PubMedID: 25596118, DOI: 10.1099/jmm.0.000021
  • Aongart Mahittikorn, Hirotake Mori, Supaluk Popruk, Amonrattana Roobthaisong, Chantira Sutthikornchai, Khuanchai Koompapong, Sukhontha Siri, Yaowalark Sukthana, Duangporn Nacapunchai (2015) Development of a Rapid, Simple Method for Detecting Naegleria fowleri Visually in Water Samples by Loop-Mediated Isothermal Amplification (LAMP). PLoS One 10, e0120997.PubMedID: 25822175, DOI: 10.1371/journal.pone.0120997
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.
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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