WarmStart® LAMP Kit (DNA & RNA) |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

WarmStart® LAMP Kit (DNA & RNA) |

The WarmStart LAMP Kit (DNA & RNA) is designed to provide a simple, one-step solution for Loop-Mediated Isothermal Amplification (LAMP) of DNA or RNA (RT-LAMP) targets. LAMP and RT-LAMP are commonly used isothermal amplification techniques that provides 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 LAMP 2X Master Mix, which contains 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. A fluorescent dye is also supplied to enable real-time fluorescence measurement of LAMP. The WarmStart LAMP Kit is compatible with multiple detection methods, including turbidity detection, real-time fluorescence detection (when used with LAMP fluorescent dye) and end-point visualization.

The WarmStart LAMP Kit (DNA & RNA) is compatible with multiple detection methods

WarmStart® LAMP Kit (DNA & RNA) |
The WarmStart LAMP Kit (DNA & RNA) includes fluorescent dye for real-time fluorescence measurement. Alternately, detection can be accomplished by turbidity detection or a variety of endpoint visualization methods.

For carryover contamination prevention, a version containing dUTP and thermolabile UDG is available: WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) (NEB #E1708).


The WarmStart LAMP Kit (DNA & RNA) offers speed and robust sensitivity

WarmStart® LAMP Kit (DNA & RNA) |
A RNA target (HMBS2) was amplified from Jurkat total RNA using the WarmStart LAMP Kit and OptiGene Master Mix (ISO-001). Reactions were performed at 65°C for 74 minutes on a real-time thermocycler (Bio-Rad CFX96) in triplicate. Time to result is set as the time at which the fluorescence crossed a threshold of 10% of maximal fluorescence. NEB’s WarmStart LAMP Kit resulted in faster and more robust detection as compared to the OptiGene Master Mix.


The WarmStart LAMP Kit (DNA & RNA) provides fast LAMP/RT-LAMP results

WarmStart® LAMP Kit (DNA & RNA) |
Experiments were designed to amplify a DNA target (BRCAb) and a RNA target (HMBS2) by LAMP or RT-LAMP, respectively. A 25 μl reaction containing 10 ng HeLa DNA or 5 ng Jurkat total RNA, 1X Master Mix, 1X LAMP primers and fluorescent dye was incubated on a Bio-Rad CFX98 instrument at 65°C for 75 minutes. Lucigen® OmniAmp® RNA & DNA Kit and OptiGene Isothermal Master Mix (ISO-001 ) were run according to manufacturers’ recommendations.


The WarmStart LAMP Kit (DNA & RNA) enables specific amplification across multiple species/targets

PRIMER SET

DNA TEMPLATE

HeLa 
DNA

Lyme 
DNA

B. malayi 
DNA

E. coli
DNA

C. elegans 
DNA

BRCAb (human)

+

 

 

 

 

CFTR (human)

+

 

 

 

 

LymO (Lyme)

 

+

 

 

 

BmHhal (B. malayi)

 

 

+

 

 

dnaE (E. coli)

 

 

 

+

 

Lec10b (C. elegans)

 

 

 

 

+

Lec6 (C. elegans)

 

 

 

 

+

In each individual reaction, multiple gDNA templates were added along with primers specific to a target in only one species. Reactions were incubated at 65°C for 74 minutes on a thermocycler (Bio-Rad CFX96) in triplicate. Positive results (+) were observed only in the expected species/target combinations, demonstrating high specificity of the WarmStart LAMP reactions.

产品类别:
Isothermal Amplification & Strand Displacement Products

应用:
Loop-Mediated Isothermal Amplification,
Isothermal Amplification

  • 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
    • E1700S     -20    
        WarmStart LAMP 2X Master Mix M1700SVIAL -20 1 x 1.25 ml 2 X
        LAMP Fluorescent Dye B1700AVIAL -20 1 x 0.05 ml 50 X
    • E1700L     -20    
        WarmStart LAMP 2X Master Mix M1700SVIAL -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 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

    • LAMP 荧光染料
    • Bst 2.0 WarmStart DNA Polymerase
    • m0380-warmstart-rtx-reverse-transcriptase
    • 等温扩增缓冲液套装
    • dNTP 混合液
    • m0374-bst-3-0-dna-polymerase

  • 参考文献

    1. Notomi, T., et al. (2000). Nucleic Acids Res. 28, PubMedID: E63
    2. Nagamine, K., et al. (2001). Clin. Chem. 47, 1742-1743.
    3. Nagamine, K., Hase, T., Notomi, T. (2002). Mol. Cell Probes. 16, 223-229.
    4. Mori, Y., Nagamine, K., Tomita, N., Notomi, T. (2001). Biochem. Biophys. Res. Commun. 289, 150-154.
    5. Mori, Y., Kitao, M., Tomita, N., Notomi, T. (2004). J. Biochem. Biophys. Methods. 59, 145-157.
    6. Tomita, N., Mori, Y., Kanda, H., Notomi, T. (2008). Nat. Protoc. 3, 877-882.
    7. Tanner, N.A. and Evans, T. C. Jr (2014). Curr. Protoc. Mol. Biol. 105, Unit 15.14.
    8. Hsieh et al. (2014). Chem. Comm. 50, 3747-3749.
    9. Tanner, N.A., Zhang, Y. and Evans, T.C. Jr (2015). Biotechniques. 58, 59-68.

操作说明、说明书 & 用法

  • 操作说明

    1. WarmStart LAMP Kit (DNA & RNA) Protocol (E1700)

  • 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。

    • manualE1700

  • 应用实例

    • Optimized Integration of New England Biolabs® Loop-mediated Isothermal Amplification (LAMP) Reagents with Axxin ISO Instruments

FAQs & 问题解决指南

  • FAQs

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