Product Class: Kit

WarmStart® LAMP Kit (DNA & RNA)

Product Introduction

  • Simple, one-step solution for Loop-Mediated Isothermal Amplification (LAMP) of DNA or RNA (RT-LAMP) targets
  • Compatible with multiple detection methods
  • WarmStart® technology inhibits activity at room temperature
Bioz Badge Exists : True
Catalog # Size Concentration
E1700S 100 reactions 0 %
E1700L 500 reactions 0 %

Product Information

Description

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.

NEB’s WarmStart LAMP Kit (DNA & RNA) is compatible with multiple detection methods


The NEB WarmStart LAMP Kit (DNA & RNA) includes separate fluorescent dye for real-time fluorescence measurement. Alternately, detection can be accomplished by turbidity detection or endpoint visualization.


NEB’s WarmStart LAMP Kit (DNA & RNA) offers speed and robust sensitivity


speed and sensitivity

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.


NEB’s WarmStart LAMP Kit (DNA & RNA) provides fast LAMP/RT-LAMP results


Fast LAMP/RT-LAMP results

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.

NEB 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.

Kit Components

The following reagents are supplied with this product:

Store at (°C) Concentration
LAMP Fluorescent Dye (50X) -20
WarmStart LAMP 2X Master Mix -20
Product Categories:
Isothermal Amplification & Strand Displacement Products
Applications:
Loop-Mediated Isothermal Amplification,
Isothermal Amplification

Properties & Usage

Materials Required but not Supplied

  • LAMP Primers (we recommend using PrimerExplorer for design of optimal LAMP Primers. PrimerExplorer V5 is recommended, https://primerexplorer.jp)
  • 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

Storage Temperature

-20°C

References

  1. Notomi, T., et al. (2000). Nucleic Acids Res. 28,
  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.

Protocols, Manuals & Usage

Protocols

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

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Application Notes

FAQs & Troubleshooting

FAQs

  1. What components are included in the kit, and which typical LAMP detection methods can be used?
  2. What is the optimal LAMP amplification temperature using this kit? 
  3. What is the Mg++ concentration in the WarmStart LAMP Master Mix? 
  4. How do I use the fluorescent dye?
  5. What DNA or RNA preparation method is required?
  6. How fast should I expect a result?
  7. How do I confirm a positive result?
  8. Can I set up my reaction at room temperature?
  9. What advantages does LAMP offer over PCR?
  10. How can I see the products of a LAMP reaction?
  11. Can I use dUTP and UDG for carryover contamination prevention with the WarmStart LAMP Master Mix?
  12. The WarmStart Colorimetric LAMP Master Mix has some precipitation in the tube after thawing, is this a problem?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

Licenses

Licensed under U.S Patent Nos. 7,374,913; 7,851,186; 7,846,695 including all foreign counterparts thereof and Patent Nos. EP 1,275,715; EP2,253,717; JP 4,726,381; ZL 01810370.7; BR 0015382. The purchase of this product conveys to the purchaser the limited, non-transferable right to use the purchased product to perform loop-mediated amplification (“LAMP”) and reverse transcription loop-mediated amplification (“RT-LAMP”) for research use only. LAMP is a patented technology belonging to Eiken Chemical Co., Ltd and any other use other than research may require a license from Eiken Chemical Co., Ltd. Notice to Purchaser: Nucleic acid-based aptamers for use with thermophilic DNA polymerases and Reverse Transcriptase are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. (US Patent Nos. 7,947,447; 8,404,830; 8,975,388 and their foreign counterparts). Commercial use of the aptamer-based WarmStart® LAMP Kit (DNA & RNA) may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact [email protected].