Product Class: Other

Bst 2.0 WarmStart® DNA Polymerase
icon_cloned icon_recomb icon_NEBU incubation temp heat inactivation icon_hotstart
Item# Description List Price Web Price Qty
M0538L Bst 2.0 WarmStart DNA Polymerase - 8,000 units $435.00
$391.50
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M0538M Bst 2.0 WarmStart DNA Polymerase (120,000 units/ml) - 8,000 units $435.00
$391.50
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M0538S Bst 2.0 WarmStart DNA Polymerase - 1,600 units $109.00
$98.10
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

    Product Introduction

    Optimized for Loop-Mediated Isothermal Amplification (LAMP)

    • Consistent amplification performance with the convenience of room temperature setup
    • WarmStart® technology eliminates off-target amplifilcation and offers increased reaction efficiency
    • Optimal reaction performance from 60-72 °C

    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.

    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.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    Isothermal Amplification Buffer Pack -20 10X
    Magnesium Sulfate (MgSO4) Solution -20 100 mM
    Product Categories:
    Isothermal Amplification & Strand Displacement Products
    Applications:
    Whole Genome Amplification,
    Isothermal Amplification

    Advantages and Features

    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

    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 Temperature

      -20°C

      Storage Conditions

      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 min

      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.

      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)

      Datacards

      The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)

      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?

      Troubleshooting

      • PCR Troubleshooting Guide

      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

      This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

      While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

      For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

      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.

      Licenses

      Notice to Purchaser: Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. (See Patent Nos. 5,475,096; 5,670,637; 5,696,249; 5,874,557; and 5,693,502). New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the aptamer-based Bst 2.0 WarmStart™ DNA Polymerase for RESEARCH PURPOSES ONLY. Commercial use of the aptamer-based Bst 2.0 WarmStart™ DNA Polymerase requires a license from New England Biolabs, Inc. Please contact [email protected] for more information.
      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.
      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.