Bst 3.0 DNA Polymerase |NEB酶试剂 New England Biolabs

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产品信息

Bst 3.0 DNA Polymerase |

Bst 3.0 DNA Polymerase is an in silico designed homologue of Bacillus stearothermophilus DNA Polymerase I, Large Fragment engineered and fused to a novel nucleic acid binding domain for improved isothermal amplification performance and increased reverse transcription activity. Bst 3.0 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. Bst 3.0 DNA Polymerase demonstrates robust performance even in high concentrations of amplification inhibitors, including dUTP and features significantly increased reverse transcriptase activity compared to Bst DNA Polymerase.

An example of the utility and speed offered by Bst 3.0 DNA Polymerase can be seen in this publication that explores the development of a novel digital LAMP assay: Rapid pathogen-specific phenotypic antibiotic susceptibility testing using digital LAMP quantification in clinical samples.

Bst 3.0 DNA Polymerase |
Fast, single-enzyme RT-LAMP can be performed using Bst 3.0 
RT-LAMP was performed using indicated DNA polymerase and Jurkat total RNA and primers for two genes (ACTB, left; HMBS2, right). Fastest results were observed with a 2-enzyme system, Bst 2.0 and WarmStart RTx, but robust amplification was also observed using Bst 3.0 without additional RT. Bst LF, Bst 2.0 and competitor enzymes showed highly variable performance, with slow threshold times or reaction failure on one of the two targets.

产品来源

An E. coli strain that carries the engineered Bst 3.0 gene.

产品类别:
Isothermal Amplification & Strand Displacement Products

应用:
Whole Genome Amplification,
Loop-Mediated Isothermal Amplification,
Isothermal Amplification

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0374S     -20    
        Bst 3.0 DNA Polymerase M0374SVIAL -20 1 x 0.2 ml 8,000 units/ml
        Isothermal Amplification Buffer II Pack B0374SVIAL -20 1 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
    • M0374L     -20    
        Bst 3.0 DNA Polymerase M0374LVIAL -20 1 x 1 ml 8,000 units/ml
        Isothermal Amplification Buffer II Pack B0374SVIAL -20 2 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
    • M0374M     -20    
        Bst 3.0 DNA Polymerase M0374MVIAL -20 1 x 0.067 ml 120,000 units/ml
        Isothermal Amplification Buffer II Pack B0374SVIAL -20 2 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM

  • 特性和用法

    单位定义

    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.

    反应条件

    1X Isothermal Amplification Buffer II Pack
    Incubate at 65°C

    1X Isothermal Amplification Buffer II Pack
    20 mM Tris-HCl
    10 mM (NH4)2SO4
    150 mM KCl
    2 mM MgSO4
    0.1% Tween® 20
    (pH 8.8 @ 25°C)

    在不同缓冲液中的活性

    rCutSmart™ Buffer: 100%
    NEBuffer™ r1.1: 10%
    NEBuffer™ r2.1: 100%
    NEBuffer™ r3.1: 100%

    贮存溶液

    100 mM KCl
    10 mM Tris-HCl
    0.1 mM EDTA
    1 mM DTT
    0.1% Triton® X-100
    50% Glycerol
    pH 7.4 @ 25°C

    热失活

    80°C for 5 minutes

    单位活性检测条件

    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.

  • 优势和特性

    应用特性

    • Isothermal amplification (LAMP, RT-LAMP)
    • Applications requiring strand-displacement DNA synthesis
    • Reverse transcription at elevated temperature (to 72°C)
    • Amplification in the presence of impure samples or amplification inhibitors

  • 相关产品

    相关产品

    • m0380-warmstart-rtx-reverse-transcriptase
    • Magnesium Sulfate (MgSO4) Solution
    • 等温扩增缓冲液套装 II
    • dNTP 套装
    • dNTP 混合液

  • 注意事项

    1. Bst 3.0 DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
    2. Reaction temperatures above 72°C are not recommended.
    3. Bst 3.0 DNA Polymerase cannot be used for PCR or reactions requiring thermal denaturation.

操作说明、说明书 & 用法

  • 操作说明

    1. Typical LAMP Protocol (M0374)

工具 & 资源

  • Web 工具

    • NEB LAMP Primer Design Tool

FAQs & 问题解决指南

  • FAQs

    1. What is the difference between Bst DNA Polymerase, Large Fragment, Bst 2.0, and Bst 3.0 DNA Polymerase?
    2. Why would I use Bst 3.0 DNA Polymerase?
    3. Can Bst 3.0 DNA Polymerase be used in other NEBuffers?
    4. How active is Bst 3.0 at other temperatures?
    5. Does Bst 3.0 DNA polymerase incorporate dUTP?
    6. Does Bst 3.0 DNA polymerase have reverse transcriptase activity?
    7. Can Bst 3.0 DNA Polymerase be used to blunt DNA?
    8. Can Bst 3.0 DNA Polymerase be used to fill in 3′ overhangs?
    9. Can Bst 3.0 DNA Polymerase be used to remove 5′ overhangs?
    10. Can Bst 3.0 DNA Polymerase be heat inactivated?
    11. Are NEB DNA Polymerases supplied with dNTPs?
    12. What are the main causes of reaction failure using Bst 3.0 DNA Polymerase?
    13. Does Bst 3.0 DNA Polymerase have an active 3’→5′ proofreading exonuclease?
    14. Can Bst 3.0 DNA Polymerase be used in applications requiring thermal cycling?
    15. Can Bst 3.0 DNA Polymerase initiate at a nick in the DNA?
    16. Can Bst 3.0 DNA Polymerase be used in labeling reactions and partial fill in reactions?
    17. Can Bst 3.0 DNA Polymerase be diluted?
    18. When I thaw Isothermal Amplification Buffer II I see a lot of white precipitate, is this normal?
    19. When should Bst 3.0 DNA Polymerase be the enzyme of choice?
    20. How do I reduce non-template amplification (NTC) in LAMP reactions with Bst 3.0?
    21. How do I use Antarctic Thermolabile UDG for carryover prevention in LAMP reactions?
    22. Does NEB have a master mix for LAMP or RT-LAMP reactions?
    23. What is LAMP and RT-LAMP?