Product Class: Other

Bst 2.0 WarmStart® DNA Polymerase (Glycerol-free)
80 Heat

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


Catalog #M0402

Product Introduction

  • Glycerol-free version of Bst 2.0 WarmStart DNA Polymerase (NEB #M0538) to support lyophilization and automation workflows
  • Consistent amplification performance with the convenience of room temperature setup
  • WarmStart technology eliminates off-target amplification and offers increased reaction efficiency
  • Higher salt tolerance and thermostability compared to wild-type Bst DNA Polymerase, Large Fragment
  • Optimal reaction performance from 60-72°C
  • One of several products available glycerol-free
  • Also available: lyophilized LAMP/RT-LAMP mix

Product Information

Description

Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) is formulated without glycerol to support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows.

Bst 2.0 WarmStart DNA Polymerase is an in silico designed homolog 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. At temperatures above 45°C, the aptamer rapidly releases from the enzyme with no special activation step 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.

This product is a core component of the LyoPrime WarmStart® Fluorescent LAMP/RT-LAMP Mix (with UDG), a lyophilized LAMP mix offering ambient/room temperature storage and shipping and streamlined assay development. To learn about the lyophilization process, please visit www.neb.com/lyoprime.

Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) offers the same robust detection of human DNA targets in LAMP assays as the glycerol-containing enzyme.

M0402 DNA Targets

LAMP (DNA targets) experiments were performed with NEB #M0538: Bst 2.0 WarmStart DNA Polymerase or NEB #M0402: Bst 2.0 WarmStart DNA Polymerase (Glycerol-free). Reactions containing 1X LAMP primers and 0.5X LAMP Fluorescent dye were set up in quadruplicate over three logs of total Jurkat DNA (10 ng to 0.1 ng) in 96-well, 25 µl reactions. Control reactions without template (NTC) were also evaluated. Reactions were incubated at 65°C for 40 minutes and fluorescence was monitored every 15 seconds in the SYBR/FAM channel of a real-time thermocycler (Bio-Rad® CFX96). Each dot represents the time at which the fluorescence signal for a single reaction crosses the instrument-defined threshold. All four replicates were detected at each template input unless otherwise indicated (note that dots frequently overlap given similar detection time for the replicates). Overall, similar performance was observed for both glycerol-containing (NEB #M0538) and glycerol-free (NEB #M0402) enzymes at each template input. No amplification was observed in any of the no template control reactions.



This product is related to the following categories:
Isothermal Amplification & Strand Displacement Products
This product can be used in the following applications:
Isothermal Amplification,
DNA Amplification, PCR & qPCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0402L     -80    
      Bst 2.0 WarmStart® DNA Polymerase (Glycerol-free) M0402LVIAL -80 1 x 0.067 ml 120,000 units/ml
      10X Isothermal Amplification Buffer (Lyo-compatible) B1714SVIAL -20 2 x 1.25 ml 10 X

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.

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
0.1% Triton® X-100
pH 7.1 @ 25°C

Heat Inactivation

80°C for 20 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 rAlbumin.

Protocols, Manuals & Usage

Protocols

  1. Typical LAMP Protocol (NEB #M0402)

Tools & Resources

Web Tools

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 2.0® DNA Polymerase be heat inactivated?
  4. Are NEB DNA Polymerases supplied with dNTPs?
  5. Can Bst 2.0® DNA Polymerase be used in applications requiring thermal cycling?
  6. Can Bst 2.0® DNA Polymerase initiate at a nick in the DNA?
  7. When should Bst 2.0® DNA Polymerase be the enzyme of choice?
  8. Can Bst 2.0® DNA Polymerase be used at temperatures other than 65°C?
  9. Does Bst 2.0® DNA polymerase incorporate dUTP?
  10. How do I use Antarctic Thermolabile UDG for carryover prevention in LAMP reactions?
  11. What is LAMP and RT-LAMP?
  12. What are Hot Start and WarmStart® polymerases and when would I use them?
  13. What is the difference between buffers B1714: 10X Isothermal Amplification Buffer (Lyo-compatible) and B0537: 10X Isothermal Amplification Buffer?
  14. What is the Mg2+ concentration in the 10X Isothermal Amplification Buffer (Lyo-compatible)?
  15. Does B1714: 10X Isothermal Amplification Buffer (Lyo-compatible) include any excipients?
  16. Can Bst 2.0 WarmStart® DNA Polymerase (Glycerol-free) at 120,000 U/ml be diluted prior to use?
  17. How many Freeze/Thaw cycles can Bst 2.0 WarmStart® DNA Polymerase (Glycerol-free) tolerate?

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.

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