Product Class: Other

Bst-XT WarmStart™ DNA Polymerase

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

This product is available in a glycerol-free format.

 


Catalog #M9204

Product Introduction

  • Combines the high specificity of Bst 2.0 and the polymerization speed of Bst 3.0 DNA Polymerases
  • Fused to novel nucleic acid binding domain for enhanced performance
  • Consistent amplification performance with the convenience of room temperature setup
  • WarmStart® technology eliminates off-target amplification and offers increased reaction efficiency
  • Offers fast polymerization rates and robust performance even in the presence of inhibitors, including dUTP
  • Broad optimal reaction performance from 50-70°C
  • View the Bst-XT: An improved Bst DNA polymerase to support loop mediated isothermal amplification NEB TV webinar
  • One of several products available glycerol-free

Product Information

Description

Bst-XT WarmStart™ DNA Polymerase is an engineered variant of Bacillus stearothermophilus DNA Polymerase, Large Fragment fused to a novel nucleic acid binding domain for improved isothermal amplification performance. This enzyme combines the high specificity of Bst 2.0 and the fast polymerization speed of Bst 3.0 DNA polymerases.

  Bst-XT WarmStart
NEB #M9204		 Bst 2.0 WarmStart
NEB #M0538		 Bst 3.0
NEB #M0374
Icon for Bst polymerase product selection Amplification Speed ★★★★★ ★★★ ★★★★★
Icon for Bst polymerase product selection Specificity ★★★★★ ★★★★★ ★★
Icon for Bst polymerase product selection Room temp. set-up? Enabled Enabled Not recommended
Icon for Bst polymerase product selection Optimal LAMP temp 50-70°C 60-70°C 55-72°C
  Icon for Bst polymerase product selection Available glycerol-free Yes
NEB #M9205		 Yes
NEB #M0402		 Yes
NEB #M0443

Bst-XT WarmStart DNA Polymerase contains a reversibly bound aptamer, which inhibits polymerase activity at room temperature. The aptamer rapidly releases above 45°C, therefore no separate activation step is needed.

Bst-XT WarmStart 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.

To support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows, Bst-XT WarmStart DNA Polymerase is also offered in a glycerol-free format.

Figure 1: Bst-XT WarmStart™ DNA Polymerase combines the high specificity of Bst 2.0 WarmStart® DNA Polymerase with the fast amplification speed of Bst 3.0 DNA Polymerase

Plots of DNA and RNA targets comparing Bst-XT WarmStart with Bst 2.0 WarmStart and Bst 3.0 WarmStart

LAMP (DNA targets, left panel) or RT-LAMP (RNA targets, right panel) experiments were performed with Bst 2.0 WarmStart DNA Polymerase (NEB #M0538), Bst 3.0 DNA Polymerase (NEB #M0374), and Bst-XT WarmStart™ DNA Polymerase (NEB #M9204) in their respective isothermal amplification buffers. To detect RNA targets, WarmStart RTx Reverse Transcriptase (NEB #M0380) was included in testing. Reactions containing 1X LAMP primers and 0.5X LAMP Fluorescent Dye (NEB #B1700) were set up in triplicate over three logs of total human Jurkat total RNA or Jurkat gDNA (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 30 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 three replicates were detected at each template input unless otherwise indicated (note that dots frequently overlap given similar detection time for the replicates). Overall, Bst-XT WarmStart DNA Polymerase showed similar time to detection for all template inputs to Bst 3.0 but without amplification in the no template controls. This data highlights that Bst-XT WarmStart DNA Polymerase combines the specificity of Bst 2.0 WarmStart DNA polymerase and the amplification speed of Bst 3.0 DNA Polymerase.

 

Figure 2: Bst-XT WarmStart™ DNA Polymerase can be used to perform LAMP reactions across a broader range of temperatures than either Bst 2.0 WarmStart® DNA Polymerase or Bst 3.0 DNA polymerase

Graph of Average Time versus Reaction Temperature, showing the ranges of Bst-XT WarmStart, Bst 2.0 WarmStart and Bst 3.0 WarmStart

LAMP (DNA target VEGFA) experiment was performed using Bst 2.0 WarmStart DNA Polymerase (NEB #M0538), Bst 3.0 DNA Polymerase (NEB #M0374), and Bst-XT WarmStart DNA Polymerase (NEB #M9204) in their respective isothermal amplification buffers. Reactions containing 1X LAMP primers and 0.5X LAMP Fluorescent Dye (NEB #B1700) were set up using Jurkat gDNA at 1 ng input in 96-well, 25 µl reactions. Reactions were incubated at temperatures ranging between 50-74°C for 60 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 average of three replicates for time to detection, which is the time at which the fluorescence signal crosses the instrument-defined threshold. Bst-XT WarmStart DNA Polymerase was able to detect 1 ng of target within 30 minutes at reaction temperatures ranging from 50°C-70°C. Bst 2.0 WarmStart DNA Polymerase showed optimal detection at temperatures between 60°C-70°C, while the optimum temperature for Bst 3.0 DNA polymerase ranged from 55°C-72°C. This data highlights that Bst-XT WarmStart DNA Polymerase can be used to perform LAMP reactions across a broader range of temperatures than either Bst 2.0 WarmStart DNA Polymerase or Bst 3.0 DNA polymerase.
This product is related to the following categories:
Isothermal Amplification & Strand Displacement
This product can be used in the following applications:
Isothermal Amplification,
DNA Amplification, PCR & qPCR

Properties & Usage

Storage Buffer

20 mM Tris-HCl
100 mM KCl
0.5 mM TCEP
0.1 mM EDTA
1X stabilizers
50% Glycerol
pH 7.5

Protocols, Manuals & Usage

Protocols

  1. Loop-mediated Isothermal Amplification (LAMP) Protocol using Bst-XT WarmStart™ DNA Polymerase (NEB #M9204)

Application Notes

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. Can Bst-XT DNA Polymerase be heat inactivated?
  2. Can Bst-XT DNA Polymerase be used at temperatures other than 65°C for LAMP reactions?
  3. Can Bst-XT DNA Polymerase be diluted?
  4. Can Bst-XT DNA Polymerase be used in end point colorimetric detection?
  5. Can Bst-XT DNA Polymerase be used with other NEB buffers?
  6. How can I optimize LAMP/RT-LAMP reactions for difficult targets when using Bst-XT?
  7. How do I use Antarctic Thermolabile UDG for carryover prevention in LAMP reactions?
  8. What are Hot Start and WarmStart® polymerases and when would I use them?
  9. What is LAMP and RT-LAMP?
  10. What is the difference between Bst DNA Polymerase, Large Fragment, Bst 2.0, Bst 3.0 and Bst-XT DNA Polymerase?
  11. What is the recommended concentration of Mg+2 to use with Bst-XT in LAMP reactions?

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.


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]

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 [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.

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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