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

#E1708

Catalog # Size Concentration Price (£)
E1708L 500 REACTIONS 2 X £1005.00
E1708S 100 REACTIONS 2 X £251.00

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  • 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

 

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



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



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

 
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.


Kit Components

The following reagents are supplied with this product:

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The following link is to a PDF Safety Data Sheet (SDS) that applies to this product to help you use it safely.



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