Bst 2.0 WarmStart DNA Polymerase |NEB酶试剂 New England Biolabs

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产品信息

Bst 2.0 WarmStart DNA Polymerase |

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.

To support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows, Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) is also offered in a glycerol-free format.

Benefits of Bst 2.0 WarmStart Bst 2.0 WarmStart DNA Polymerase |
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.

产品来源

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.

产品类别:
Isothermal Amplification & Strand Displacement Products

应用:
Whole Genome Amplification,
Strand Displacement Amplification & Nicking Enzyme,
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
    • M0538S     -20    
        Bst 2.0 WarmStart® DNA Polymerase M0538SVIAL -20 1 x 0.2 ml 8,000 units/ml
        Isothermal Amplification Buffer Pack B0537SVIAL -20 1 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
    • M0538L     -20    
        Bst 2.0 WarmStart® DNA Polymerase M0538LVIAL -20 1 x 1 ml 8,000 units/ml
        Isothermal Amplification Buffer Pack B0537SVIAL -20 2 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
    • M0538M     -20    
        Bst 2.0 WarmStart® DNA Polymerase M0538MVIAL -20 1 x 0.067 ml 120,000 units/ml
        Isothermal Amplification Buffer Pack B0537SVIAL -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 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)

    贮存溶液

    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

    热失活

    80°C for 20 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)
    • Applications requiring strand-displacement DNA synthesis
    • DNA sequencing through high GC regions
    • Rapid sequencing from nanogram amounts of DNA template

  • 相关产品

    相关产品

    • m0537-bst-20-dna-polymerase
    • m0275-bst-dna-polymerase-large-fragment
    • dNTP 混合液
    • dNTP 套装
    • 等温扩增缓冲液套装
    • Magnesium Sulfate (MgSO4) Solution

  • 注意事项

    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.

操作说明、说明书 & 用法

  • 操作说明

    1. Typical LAMP Protocol (M0538)
    2. Strand Displacement Amplification (SDA) Protocol using WarmStart® Nt.BstNBI (NEB #R0725)

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 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?
    20. What is LAMP and RT-LAMP?
    21. What are Hot Start and WarmStart® polymerases and when would I use them?

  • 问题解决指南

    • PCR Troubleshooting Guide