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

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Bst DNA Polymerase, Large Fragment

The original polymerase for Loop-Mediated Isothermal Amplification (LAMP)

  • Modified to retain 5´ → 3´ polymerase activity while lacking 5´ →3´ exonuclease activity
  • Suitable for applications requiring thermophilic strand displacement
  • Supplied with ThermoPol® Reaction Buffer for high product yields in demanding conditions

Bst DNA Polymerase, Large Fragment |

库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
M0275V 8,000 units/ml 800 units ¥409.00 有 有 无 有 有
M0275S 8,000 units/ml 1,600 units ¥759.00 有 有 有 有 有
M0275L 8,000 units/ml 8,000 units ¥3,049.00 有 有 有 无 有
M0275M 120,000 units/ml 8,000 units ¥3,049.00 无 无 有 无 无
如何订购
Please enter a quantity for at least one size

Bst DNA Polymerase, Large Fragment | Interested in glycerol-free or custom formulations? – contact our Custom Solutions Group

Bulk packaging may also be available and requested for large recurring orders.
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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

    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 Pack 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 Pack 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 Pack 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 Pack 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 Pack
    Incubate at 65°C

    1X ThermoPol® Reaction Buffer Pack
    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 min

    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

    • Deoxynucleotide (dNTP) Solution Set
    • Magnesium Sulfate (MgSO4) Solution
    • Deoxynucleotide Solution Mix
    • Bst 2.0 DNA Polymerase
    • Bst 2.0 WarmStart DNA Polymerase

    单独销售的组分

    • ThermoPol® Reaction Buffer Pack

  • 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?

  • 问题解决指南
    • PCR Troubleshooting Guide

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    Bst DNA Polymerase, Large Fragment | Powered by Bioz See more details on Bioz

    更多引用文献

    • Trisadee Khamlor, Petai Pongpiachan, Rangsun Parnpai, Kanchana Punyawai, Siwat Sangsritavong, Nipa Chokesajjawatee (2015) Bovine embryo sex determination by multiplex loop-mediated isothermal amplification. Theriogenology; 83, 891-6. PubMedID: 25542460, DOI: 10.1016/j.theriogenology.2014.11.025
    • DoKyung Lee, Eun Jin Kim, Paul E Kilgore, Soon Ae Kim, Hideyuki Takahashi, Makoto Ohnishi, Dang Duc Anh, Bai Qing Dong, Jung Soo Kim, Jun Tomono, Shigehiko Miyamoto, Tsugunori Notomi, Dong Wook Kim, Mitsuko Seki (2015) Clinical Evaluation of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Neisseria meningitidis in Cerebrospinal Fluid. PLoS One; 10, e0122922. PubMedID: 25853422, DOI: 10.1371/journal.pone.0122922
    • Manabu Nemoto, Yoshinori Morita, Hidekazu Niwa, Hiroshi Bannai, Koji Tsujimura, Takashi Yamanaka, Takashi Kondo (2015) Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods; 215-216, 13-6. PubMedID: 25682750, DOI: 10.1016/j.jviromet.2015.02.001
    • Mohammad Reza Allahyar Torkaman, Kazunari Kamachi, Vajihe Sadat Nikbin, Masoumeh Nakhost Lotfi, Fereshteh Shahcheraghi (2015) Comparison of loop-mediated isothermal amplification and real-time PCR for detecting Bordetella pertussis. J Med Microbiol; 64, 463-5. PubMedID: 25596118, DOI: 10.1099/jmm.0.000021
    • Aongart Mahittikorn, Hirotake Mori, Supaluk Popruk, Amonrattana Roobthaisong, Chantira Sutthikornchai, Khuanchai Koompapong, Sukhontha Siri, Yaowalark Sukthana, Duangporn Nacapunchai (2015) Development of a Rapid, Simple Method for Detecting Naegleria fowleri Visually in Water Samples by Loop-Mediated Isothermal Amplification (LAMP). PLoS One; 10, e0120997. PubMedID: 25822175, DOI: 10.1371/journal.pone.0120997

质控、安全 & 法规

  • 质控分析
    每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。

  • 产品说明与变更通知

    产品说明与变更通知

    产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]

    • M0275M_v1
    • M0275S_L_v1
    • M0275M_v2
    • M0275S_L_v2

  • CoA
    CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]

    • M0275S_L_v1_0511509
    • M0275M_v1_0511509
    • M0275M_v1_0511603
    • M0275S_L_v1_0511603
    • M0275M_v1_0511609
    • M0275S_L_v1_0511609
    • M0275S_L_v1_0511703
    • M0275M_v1_0511703
    • M0275S_L_v1_0511709
    • M0275M_v1_0511709
    • M0275M_v1_0511803
    • M0275S_L_v1_0511803
    • M0275L_v1_10011610
    • M0275S_v1_10009342
    • M0275M_v1_10017700
    • M0275L_v1_10020437
    • M0275S_v1_10032197
    • M0275L_v1_10029594
    • M0275S_v1_10038935
    • M0275M_v1_10035567
    • M0275L_v1_10046214
    • M0275S_v1_10045900
    • M0275S_v1_10048185
    • M0275L_v1_10051813
    • M0275M_v1_10055954
    • M0275L_v1_10053202
    • M0275S_v1_10059774
    • M0275S_v2_10063500
    • M0275L_v1_10063499
    • M0275L_v1_10067894
    • M0275M_v2_10068169
    • M0275S_v2_10068243
    • M0275L_v2_10072178
    • M0275M_v2_10073620
    • M0275S_v2_10078286
    • M0275L_v2_10075398
    • M0275M_v2_10086262
    • M0275S_v2_10088116
    • M0275S_v2_10094232
    • M0275M_v2_10095463
    • M0275L_v2_10100585
    • M0275S_v2_10108804
    • M0275S_v2_10115484
    • M0275L_v2_10108569
    • M0275M_v2_10127396
    • M0275S_v2_10122371
    • M0275L_v2_10136893
    • M0275M_v2_10140685
    • M0275S_v2_10153544
    • M0275M_v2_10160590
    • M0275M_v2_10164128
    • M0275L_v2_10162366

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Bst DNA Polymerase, Large Fragment
    • ThermoPol® Reaction Buffer Pack
    • Magnesium Sulfate (MgSO4) Solution