WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) ​ |NEB酶试剂 New England Biolabs

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WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG)

  • Simple, one-step solution for Loop-Mediated Isothermal Amplification (LAMP) of DNA or RNA (RT-LAMP) targets
  • Reduce risk of carryover contamination with thermolabile UDG and dUTP included in the master mix
  • Includes fluorescent dye to enable real-time fluorescence measurement of LAMP/RT-LAMP reaction
  • WarmStart technology inhibits activity at room temperature
  • Learn more about how RT-LAMP is being used for rapid COVID-19 screening
  • Master mix (NEB #M1708), LAMP Fluorescent Dye (NEB #B1700) components available separately
 

精选视频

WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) ​ |

NEB® LAMP Primer Design Tool Tutorial

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库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
E1708S 2 X 100 reactions ¥3,279.00 无 无 无 无 无
E1708L 2 X 500 reactions ¥13,099.00 无 无 无 无 无
如何订购
Please enter a quantity for at least one size

WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) ​ | Need a custom/large volume order? Contact us

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

The WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) is designed to provide a simple, one-step solution for real-time detection of LAMP/RT-LAMP reactions via fluorescence. LAMP and RT-LAMP are commonly used isothermal amplification techniques that provide rapid detection of a target nucleic acid using LAMP-specific primers (supplied by the user) and a strand-displacing DNA polymerase. This kit is supplied with the WarmStart Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) (NEB #M1708) featuring a blend of Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx Reverse Transcriptase in an optimized LAMP buffer solution. Both Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx Reverse Transcriptase have been engineered for improved performance in LAMP and RT-LAMP reactions. Also included is a 50X LAMP Fluorescent Dye for real-time fluorescence measurement.

The inclusion of dUTP and thermolabile UDG in the master mix reduces the possibility of carryover contamination, where unintended product of a previous amplification can serve as the substrate of a subsequent reaction. Thermolabile UDG is completely inactivated at temperatures above 50°C, thereby having no effect on the reaction.

Figure 1: LAMP/RT-LAMP master mixes and sample type considerations

WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) ​ |

NEB’s pH-based colorimetric LAMP master mixes with UDG (NEB #M1804) or without UDG (NEB# M1800) are weakly buffered to allow for visual detection of amplification using a pH-sensitive dye. However, the low buffering capacity required to generate the pink-to-yellow color change limits sample compatibility with the pH-based colorimetric mixes, as highly buffered sample inputs or acidic samples may impact the color change. The multi-purpose LAMP/RT-LAMP 2X Master Mix with UDG (NEB #M1708, NEB #E1708) or without UDG (NEB #E1700) is fully buffered and can more readily tolerate these types of sample inputs, making it compatible with various detection modes including fluorescence or other colorimetric dyes (e.g., hydroxynaphthol blue).

Figure 2: The WarmStart Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) provides robust detection of human DNA and RNA targets

WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) ​ |

RT-LAMP (RNA targets) or LAMP (DNA targets) experiments were performed with NEB #M1700: WarmStart LAMP 2X Master Mix, which is the master mix in the NEB #E1700 kit that does not contain dUTP/UDG, and NEB #M1708: WarmStart Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG). Reactions containing 1X LAMP primers and 1X LAMP Fluorescent dye were set up in quadruplicate over three logs of total Jurkat RNA or Jurkat DNA (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 40 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 four replicates were detected at each template input unless otherwise indicated (note that dots frequently overlap given similar detection time for the replicates). Overall, similar performance was observed for both NEB #M1700 and NEB #M1708 at each template input.No amplification was observed in any of the no template control reactions.

Figure 3: The WarmStart Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) is compatible with automated reaction assembly

WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) ​ | 

LAMP assays targeting synthetic SARS-CoV-2 RNA were assembled either manually or with the TEMPEST® liquid handling platform (96-well, 25 µl reactions). The assay was carried out using either positive samples (human total RNA plus synthetic SARS-CoV-2 RNA at 5,000, 500 or 50 copies per reaction) or no template (NTCs) as indicated. Reactions were incubated at 65°C for 40 minutes and monitored with 1X LAMP Fluorescent dye in the SYBR/FAM channel of a real-time instrument (Bio-Rad CFX96). Similar time to detection and LOD results were observed for both reaction assembly methods.

产品类别:
Isothermal Amplification & Strand Displacement Products

应用:
Loop-Mediated Isothermal Amplification

  • 试剂盒组成

    试剂盒组成

    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
    • E1708S     -20    
        WarmStart® Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) M1708SVIAL -20 1 x 1.25 ml 2 X
        LAMP Fluorescent Dye B1700AVIAL -20 1 x 0.05 ml 50 X
    • E1708L     -20    
        WarmStart® Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) M1708SVIAL -20 5 x 1.25 ml 2 X
        LAMP Fluorescent Dye B1700SVIAL -20 1 x 0.25 ml 50 X

  • 特性和用法

    需要但不提供的材料

    LAMP Primers (we recommend using the NEB LAMP Primer Design Tool)
    Target nucleic acid samples
    Molecular Biology Grade H2O
    Heat block, water bath, real-time turbidimeter or thermocycler (with real-time fluorescence measurement if desired) and instrument-appropriate reaction vessels
     

  • 相关产品

    相关产品

    • WarmStart® Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG)
    • LAMP Fluorescent Dye
    • Bst 2.0 WarmStart DNA Polymerase
    • WarmStart® RTx Reverse Transcriptase

操作说明、说明书 & 用法

  • 操作说明
    1. WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) Protocols (NEB #E1708)

  • 说明书
    产品说明书包含产品使用的详细信息、产品配方和质控分析。

    • manualE1708

  • 应用实例
    • Optimizing rapid isothermal workflow for SARS-CoV-2 with WarmStart® LAMP with UDG

工具 & 资源

  • Web 工具
    • NEB LAMP Primer Design Tool

FAQs & 问题解决指南

  • FAQs
    1. What is the optimal LAMP/RT-LAMP amplification temperature?
    2. What is the Mg++ concentration in the WarmStart LAMP/RT-LAMP Master Mix?
    3. How do I use the fluorescent dye?
    4. What types of input samples/materials are compatible with the LAMP/RT-LAMP mixes?
    5. How fast should I expect a result?
    6. How do I confirm a positive result?
    7. Can I set up my LAMP/RT-LAMP reactions at room temperature?
    8. The WarmStart LAMP Master Mix has some precipitation in the tube after thawing, is this normal?
    9. How do I design LAMP Primers?
    10. What is the difference between NEB #E1708: WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) and NEB #E1700: WarmStart LAMP Kit (DNA & RNA)?
    11. What components are included in the WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG)?
    12. Does the WarmStart Multi-purpose LAMP/RT-LAMP 2X Master Mix (with UDG) enable carryover prevention/contamination reduction?
    13. Amplification occurred in my NTC sample(s) following isothermal incubation. What happened?
    14. What are the advantages of using standard LAMP (NEB #E1700, M1708, E1708) over pH-based colorimetric LAMP (NEB #M1800, M1804)?
    15. Is a separate incubation step required for RT-LAMP?
    16. Does NEB have a master mix for LAMP or RT-LAMP reactions?
    17. What is LAMP and RT-LAMP?

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]

    • E1708S_L_v1
    • E1708S_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#]

    • E1708L_v1_10114221
    • E1708S_v1_10114298
    • E1708S_v2_10135812
    • E1708L_v2_10135818
    • E1708L_v2_10137402
    • E1708L_v2_10139438
    • E1708S_v2_10138396
    • E1708S_v2_10141435
    • E1708L_v2_10141014
    • E1708S_v2_10144803
    • E1708S_v2_10145386
    • E1708L_v2_10148981
    • E1708S_v2_10153100
    • E1708L_v2_10155082
    • E1708S_v2_10155831
    • E1708L_v2_10157245
    • E1708S_v2_10158366
    • E1708S_v2_10162277
    • E1708L_v2_10159701

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • WarmStart® Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG)
    • LAMP Fluorescent Dye