Product Class: Kit

SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit

This product is intended for research purposes only.

Product Introduction

The SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit utilizes isothermal amplification for use in the analysis of SARS-CoV-2, the novel coronavirus that causes COVID-19.

  • Colorimetric LAMP enables simple, visual detection (pink-to-yellow) of amplification of SARS-CoV-2 nucleic acid
  • Set up reactions quickly and easily, using a simple heat source and unique WarmStart® technology
  • Reduce risk of carryover contamination, with UDG and dUTP included in the master mix
  • Assay targets N and E regions of the SARS-CoV-2 genome, for optimized sensitivity and specificity
  • Bring confidence to your results using the provided controls
  • Learn more about LAMP and other isothermal amplification methods
  • Learn more about how NEB is supporting COVID-19 research
 
 
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Catalog # Size Concentration
E2019S 96 reactions 0

Product Information

Description

The SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit utilizes Loop-Mediated Isothermal Amplification (LAMP) to detect SARS-CoV-2 nucleic acid. The kit is available for research use only and includes WarmStart Colorimetric LAMP 2X Master Mix with UDG and a primer mix targeting the N and E regions of the viral genome. Controls are provided to verify assay performance, and include an internal control primer set and a positive control template. Guanidine hydrochloride has been found to increase the speed and sensitivity of the RT-LAMP reaction and is also included.

Figure 1: SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit Components

Get results in just 30 minutes with NEB’s SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit.

The SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit includes WarmStart Colorimetric LAMP 2X Master Mix with UDG and a primer mix targeting the N and E regions of the viral genome. Controls are provided to verify assay performance and include an internal control primer set and a positive control template. Guanidine hydrochloride has been found to increase the speed and sensitivity of the RT-LAMP reaction and is also included.



Figure 2: SARS-CoV-2 Genome and SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit Gene Targets

 See all components in NEB’s SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit

The SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit includes a dual primer mix that targets the N and E gene regions.



WarmStart Colorimetric LAMP 2X Master Mix with UDG is an optimized formulation of Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx in a special low-buffer reaction solution containing a visible pH indicator for rapid and easy detection of LAMP and RT-LAMP reactions. The inclusion of dUTP and UDG in the master mix reduces the possibility of carryover contamination between reactions.
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This system provides a fast, clear visual detection of amplification based on the production of protons and the subsequent drop in pH that occurs from the extensive DNA polymerase activity in a LAMP reaction. The decrease in pH produces a change in solution color from pink to yellow. The total protocol time is 30 minutes and only requires a simple heating device that can reach 65°C.
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A positive detection of the SARS-CoV-2 RNA sequence would be indicated by a yellow color (amplification occurred, protons released, pH-dependent color change from pink to yellow), while a negative result would be indicated by a pink color (no amplification occurred, no protons released, no color change).
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Figure 3: SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit Workflow



In the SARS-CoV-2 Colorimetric LAMP Assay Kit, three control reactions are run with each test sample. All reactions should be pink prior to incubation. Using the provided protocol, a Non-Template Control (NTC) reaction will contain all materials of the test sample (master mix, primers, etc) except for the test input nucleic acid itself and serves as a measure of reaction contamination and primer-based mis-amplification. The NTC sample should not amplify and should stay pink throughout the experiment. The Positive Control (PC) will contain master mix, a plasmid that contains the SARS-CoV-2 N-gene (GenBank: MN908947.3) and primers that will amplify this sequence. Amplification should be observed in the PC and the sample should become yellow after incubation. The Internal Control (IC) will contain master mix, test input nucleic acid, and LAMP primers for rActin, an endogenous housekeeping gene. If reagents are active and samples have been handled appropriately, the IC should become yellow after incubation.


 
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Figure 4: Rapid, simple, color-based detection of SARS-CoV-2 RNA



The SARS-CoV-2 Rapid Colorimetric LAMP Assay was carried out using the indicated controls and either positive sample (human total RNA + synthetic SARS-CoV-2 RNA) or negative sample (human total RNA alone). Valid results for Non-Template Control (NTC, pink), Positive Control (PC, yellow) and Internal Control (IC, yellow) are shown. In the positive test sample, isothermal amplification of SARS-CoV-2 RNA leads to a  color change from pink to yellow.



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Figure 5: SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit Workflow



The SARS-CoV-2 Rapid Colorimetric LAMP Assay was carried out using either positive samples (human total RNA + synthetic SARS-CoV-2 RNA at 5,000, 500 or 50 copies per reaction), negative samples (human total RNA alone) or no template (NTCs) as indicated. A. All positive samples gave a positive result with robust color change from pink to yellow, including detection in all samples at a low input of 50 copies per reaction (n = 20). B. No color change was observed for negative samples (n = 32). C. Summary of results from analogous testing conducted by 3 users in 9 experiments total. At 50 copies per reaction, >95% (277/288) of reactions were positive (yellow). For negative samples, >99% (358/360) of reactions gave no color change (pink) and 2/360 reactions gave a partial color change (orange, as indicated by asterisk for Users 2 and 3).


LAMP has been used in novel workflows by scientists around the world to further COVID-19 research. To access application notes, selected preprints, and to learn more about NEB’s COVID-19 Researcher Spotlight series, click here.   

Kit Components

The following reagents are supplied with this product:

Store at (°C) Concentration
WarmStart® Colorimetric LAMP 2X Master Mix with UDG -20 2 X
SARS-CoV-2 Positive Control (N gene) -20 12.5 X
Internal Control LAMP Primer Mix -20 10 X
SARS-CoV-2 LAMP Primer Mix (N/E) -20 10 X
Nuclease-free Water -20
Guanidine Hydrochloride -20 10 X
Product Categories:
Isothermal Amplification & Strand Displacement Products,
PCR, qPCR & Amplification Technologies Products
Applications:
Isothermal Amplification,
DNA Amplification, PCR & qPCR

Advantages and Features

Features

  • 30-minute isothermal protocol – no sophisticated lab equipment required
  • Colorimetric, visual detection – ease of use, simple-to-read results
  • UDG and dUTP included in master mix for carryover prevention – reduced risk of contamination
  • Optimized dual primer mix – enhanced detection and sensitivity
  • Dual WarmStart formulation – room temperature setup
  • Necessary controls included – verify assay performance for increased confidence in results

Properties & Usage

Materials Required but not Supplied

Disposable powder-free gloves and any additional PPE required

P2/P10, P200, and P1000 aerosol barrier tips

Sterile, nuclease-free 1.5 ml microcentrifuge tubes

Sterile, nuclease-free 2.0 ml, 5.0 ml, or 15 ml tubes

0.2 ml PCR reaction tube strips with separated tubes and lids (e.g., VWR 20170-004) or attached caps (e.g., VWR 20170-010) or 96-well PCR reaction plates with 8-cap strips

Racks for 1.5 ml microcentrifuge tubes and 96-well 0.2 ml PCR reaction tubes

Cooler rack for 1.5 microcentrifuge tubes and 96-well 0.2 ml PCR reaction tubes

Acceptable surface decontaminants, for example: 10% bleach (1:10 dilution of commercial 5.25-6.0% sodium hypochlorite)

Laboratory marking pen

Appropriate disposal containers

White paper or light background for optimal visualization of colorimetric reaction (e.g., typical printer paper)

Micropipettes (2 or 10 μl, 200 μl and 1,000 μl), Multichannel Micropipettes (5-50 μl)

-20°C Freezer (frost-free or nonfrost), 4°C Refrigerator

Thermocycler, heat block or device that can be set to 65°C

PCR Work Station [UV lamp; Laminar flow (Class 100 HEPA filtered)], Vortex Mixer, Tabletop Microcentrifuge

Storage Temperature

-20°C

Product Notes

  1. Carryover Contamination Prevention

    LAMP is a sensitive method that generates large quantities of DNA, and contamination in new LAMP assays with products from previous amplification reactions can cause a variety of issues, such as false positive results and a decrease in sensitivity. The best way to prevent this “carryover” contamination is to practice good laboratory procedures and avoid opening the reaction vessel post amplification. However, to accommodate situations where additional anti-contamination measures are desired, WarmStart Colorimetric LAMP Master Mix with UDG contains a mixture of dUTP/dTTP that results in the incorporation of dU into the DNA product during amplification. Antarctic Thermolabile uracil DNA glycosylase (UDG) present in the colorimetric mix will eliminate previously-amplified uracil-containing products by excising the uracil base to produce a non-amplifiable DNA product. The use of a thermolabile UDG is important, as quick and complete inactivation of the UDG is required to prevent destruction of newly synthesized LAMP products. To maximize elimination of contaminating products, set up colorimetric LAMP experiments at room temperature or include a 10 minute incubation step at 25°C before isothermal incubation.

  2. Reaction Setup and Isothermal Incubation

    Due to the dual WarmStart feature of the Colorimetric LAMP Master Mix with UDG, reactions may be set up at room temperature. Room temperature set up will also allow UDG to destroy previously generated dU-containing LAMP products.

  3. We recommend a final reaction volume of 25 μl.
  4. For maximum sensitivity, go directly from room temperature to a preheated block set at 65°C. Reactions should not be allowed to sit on a block as it warms to 65°C.

Protocols, Manuals & Usage

Protocols

  1. SARS-CoV-2 Rapid Colorimetric LAMP Detection Assay Protocol (NEB #E2019)

Manuals

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

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. Can the Colorimetric LAMP 2X Master Mix be used in instruments that utilize fluorescent detection of DNA amplification or real time turbidity measurement?
  2. How stable are the WarmStart Colorimetric LAMP 2X Master Mixes?
  3. The WarmStart Colorimetric LAMP Master Mix has some precipitation in the tube after thawing, is this a problem?
  4. What if my colorimetric LAMP mix is orange instead of pink prior to amplification?
  5. Does the WarmStart® Colorimetric LAMP Master Mix with UDG enable carryover prevention/contamination reduction?
  6. Does pre-heating the thermal block impact LAMP amplification reactions?
  7. What regions of the SARS-CoV-2 virus do the LAMP Primers target? How were these primers chosen?
  8. In the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit, what is the target of the Internal Control Primer Mix (rActin)?
  9. Should I use Guanidine HCl (B2619A) in colorimetric LAMP reactions?
  10. How are colorimetric LAMP results interpreted in the SARS-CoV-2 Kit?
  11. After amplification my samples turned orange rather than yellow. Is this acceptable? How do I interpret the results?
  12. Can I incubate my colorimetric LAMP Sars-CoV-2 samples longer than 30 minutes at 65°C? How long should I incubate the reaction before checking the results?
  13. My NTC sample(s) turned orange/yellow following incubation at 65°C. What happened?
  14. My colorimetric LAMP SARS-CoV-2 Positive Control failed to turn yellow following incubation at 65°C. What happened?
  15. What is the SARS-CoV-2 Positive Control included in the kit?
  16. How much sample can be added to the colorimetric LAMP reaction as input?
  17. What types of samples/materials are compatible with the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit?
  18. My SARS-CoV-2 sample or Internal Control reaction turned orange/yellow upon sample addition. Does that mean that the sample contains nucleic acid?
  19. What instruments are compatible with the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit?
  20. Can I monitor the color change of the colorimetric LAMP reactions using a spectrophotometer?

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

This product is not available for sale in China (including Hong Kong and Macau), Japan and Taiwan. For questions, please contact [email protected].

This product is licensed under U.S Patent Nos. 9,909,168; 7,374,913; 7,851,186; 7,846,695, including all foreign counterparts thereof and Patent Nos. EP 1,275,715; EP 2,253,717; BR 0015382.  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. The purchase of this product conveys to the purchaser the limited, non-transferable right, under intellectual property owned or controlled by New England Biolabs, Inc., to use the  product to perform loop-mediated amplification (“LAMP”) and reverse transcription loop-mediated amplification (“RT-LAMP”) for investigational research and in vitro diagnostic tests for the detection of severe acute respiratory syndrome coronavirus, SARS-CoV-2. 
 
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). New England Biolabs, Inc. gives the purchaser a non-exclusive license to the aptamer- based WarmStart® Colorimetric LAMP 2X Master Mix with UDG for research and in vitro diagnostic tests for the detection of severe acute respiratory syndrome coronavirus, SARS-CoV-2.  
 
This product is not approved by the US Food and Drug Administration, or any foreign equivalent, for diagnostic use. It is the purchaser’s responsibility to ensure it has appropriate authorization for its particular use.