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.
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.
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.
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).
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.
The following reagents are supplied with this product:
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.
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.
We recommend a final reaction volume of 25 μl.
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.
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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.
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