Bst DNA Polymerase, Large Fragment |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

Bst DNA Polymerase, Large Fragment |

Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the
5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity.

重点

  • Isolated from a recombinant source
  • Sequencing through problematic secondary structures
  • Supplied with 10X Reaction Buffer

产品来源

Bst Polymerase, Large Fragment is prepared from an E. coli strain containing a genetic fusion of the Bacillus stearothermophilus DNA Polymerase gene, lacking the 5´ → 3´ exonuclease domain, and the gene coding for E. coli maltose binding protein (MBP). The fusion protein is purified to near homogeneity and the MBP portion of the fusion is cleaved off in vitro. The remaining polymerase is purified free of MBP (1).

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
    • M0275V     -20    
        Bst DNA Polymerase, Large Fragment M0275VVIAL -20 1 x 0.1 ml 8,000 units/ml
        ThermoPol® Reaction Buffer B9004SVIAL -20 1 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
    • M0275S     -20    
        Bst DNA Polymerase, Large Fragment M0275SVIAL -20 1 x 0.2 ml 8,000 units/ml
        ThermoPol® Reaction Buffer B9004SVIAL -20 1 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
    • M0275L     -20    
        Bst DNA Polymerase, Large Fragment M0275LVIAL -20 1 x 1 ml 8,000 units/ml
        ThermoPol® Reaction Buffer B9004SVIAL -20 2 x 1.5 ml 10 X
        Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
    • M0275M     -20    
        Bst DNA Polymerase, Large Fragment M0275MVIAL -20 1 x 0.067 ml 120,000 units/ml
        ThermoPol® Reaction Buffer B9004SVIAL -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 at the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.

    反应条件

    1X ThermoPol® Reaction Buffer
    Incubate at 65°C

    1X ThermoPol® Reaction Buffer
    20 mM Tris-HCl
    10 mM (NH4)2SO4
    10 mM KCl
    2 mM MgSO4
    0.1% Triton® X-100
    (pH 8.8 @ 25°C)

    在不同缓冲液中的活性

    NEBuffer™ 1: 50%
    NEBuffer™ 2: 100%
    NEBuffer™ 3: 50%
    NEBuffer™ 4: 100%

    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

    分子量

    理论上的: 67000 daltons

    5′ – 3′ 核酸外切酶

    No

    3′ – 5′ 核酸外切酶

    No

    链置换

    ++++

    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)
    • DNA sequencing through high GC regions (2,3)
    • Rapid Sequencing from nanogram amounts of DNA template (4)

  • 相关产品

    相关产品

    • dNTP 套装
    • Magnesium Sulfate (MgSO4) Solution
    • Deoxynucleotide Solution Mix
    • m0537-bst-20-dna-polymerase
    • m0538-bst-20-warmstart-dna-polymerase

    单独销售的组分

    • ThermoPol® 反应缓冲液套装

  • 注意事项

    1. Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
    2. 100 µg/ml BSA or 0.1%Triton X-100 is required for long term storage.
    3. Reaction temperatures above 70°C are not recommended.
    4. Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing or PCR.

  • 参考文献

    1. Kong, H., Aliotta, J. and Pelletier, J.J. New England Biolabs. Unpublished observation
    2. Griffin, H. and Griffin, A. (1994). PCRTechnology. 228-229.
    3. McClary, J. et al. (1991). J. DNASequencing and Mapping. 1, 173-180.
    4. Mead, D.A. et al. (1991). Biotechniques. 11, 76-87.
    5. Notomi et al. (2000). Nucleic Acids Res. 28(12), E63.
    6. Hsieh et al. (2014). Chem. Commun. 50, 3747-3749.
    7. Tanner and Evans (2014). Curr. Prot. Mol. Biol. 105, 15.14.

操作说明、说明书 & 用法

  • 操作说明

    1. Typical LAMP Protocol (M0275)

  • 使用指南

    • Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

工具 & 资源

  • 选择指南

    • DNA Polymerase Selection Chart

  • Web 工具

    • NEB LAMP Primer Design Tool

FAQs & 问题解决指南

  • FAQs

    1. Can Bst DNA Polymerase be used in other NEBuffers, including rCutSmart?
    2. Can Bst DNA Polymerase be used to blunt DNA?
    3. Can Bst DNA Polymerase be used to fill in 3′ overhangs?
    4. Can Bst DNA Polymerase be used to remove 5′ overhangs?
    5. Can Bst DNA Polymerase be heat inactivated?
    6. Are NEB DNA Polymerases supplied with dNTPs?
    7. What are the main causes of reaction failure using Bst DNA Polymerase?
    8. Does Bst DNA Polymerase have an active 3’→5′ proofreading exonuclease?
    9. Can Bst DNA Polymerase be used for thermal cycle sequencing?
    10. Can Bst DNA Polymerase initiate at a nick in the DNA?
    11. Can Bst DNA Polymerase be used in labeling reactions and partial fill in reactions?
    12. Can Bst DNA Polymerase be diluted?
    13. When should Bst DNA Polymerase be the enzyme of choice?
    14. Can Bst DNA Polymerase be used at temperatures other than 65°C?
    15. Does Bst DNA Polymerase, Large Fragment incorporate dUTP?
    16. Does Bst DNA polymerase have reverse transcriptase activity?
    17. Does NEB have a master mix for LAMP or RT-LAMP reactions?
    18. What is LAMP and RT-LAMP?
    19. What is the difference between Bst DNA Polymerase, Large Fragment and Bst 2.0 DNA Polymerase?

  • 问题解决指南

    • PCR Troubleshooting Guide