Product Class: Other

LyoPrime WarmStart™ Fluorescent LAMP/RT-LAMP Mix (with UDG)

View the NEB TV webinar on Lyophilizing Reagents on demand. Please contact us to customize this product.


Catalog #L4401

Product Introduction

  • Lyophilized format for ambient/room temperature shipping and storage
  • Simple, rapid rehydration (view vial protocol, view 96-well plate protocol)
  • Store at room temperature for up to 2 years prior to rehydration
  • 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 enzyme activity at room temperature

L4401L4401

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Product Information

Description

The LyoPrime WarmStart Fluorescent LAMP/RT-LAMP Mix (with UDG) is offered in a lyophilized format, allowing for ambient/room temperature shipping and storage before use. It is a simple, one-step solution for real-time fluorescent detection of LAMP/RT-LAMP reactions. LAMP and RT-LAMP are commonly used isothermal amplification techniques that provide rapid detection of a target nucleic acid using LAMP-specific primers and a strand-displacing DNA polymerase. Featuring a blend of Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx Reverse Transcriptase in an optimized buffer solution, this mix offers improved performance. LAMP Fluorescent Dye at 0.5X concentration is included to enable detection of amplification via real time fluorescence. 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.

This product is available in 2 x 50 reaction glass vials or a 96-well plate (1 reaction per well). The plate features twelve partially-attached 8-well strip tubes, allowing for use of the entire plate or partial consumption. The yellow shell frame adaptor supports the use of the plate in a variety of common real-time PCR instruments. Both formats can be customized. Additionally, should alternative detection methods be desired (CRISPR-Cas, end-point visualization such as colorimetric detection via a metal indicator (e.g., hydroxynaphthol blue.), etc.), please contact us for more information.

Figure 1: LyoPrime LAMP offers a robust, versatile LAMP/RT-LAMP option for fluorescence detection in a shelf-stable lyophilized format

LAMP Formats

The LyoPrime WarmStart Fluorescent LAMP/RT-LAMP Mix (with UDG) (NEB #L4401) offers the same versatile features as the WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) (NEB #E1708) in a lyophilized format that can be shipped and stored at room temperature. It is available in glass vials or a 96-well plate.




Figure 2: The LyoPrime WarmStart Fluorescent LAMP/RT-LAMP Mix (with UDG) provides robust detection of human DNA and RNA targets

LAMP Human Targets

RT-LAMP (RNA targets) or LAMP (DNA targets) experiments were performed with NEB #E1708: WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) using 0.5X LAMP Fluorescent Dye, and NEB #L4401: LyoPrime WarmStart Fluorescent LAMP/RT-LAMP Mix (with UDG). Reactions containing 1X LAMP primers 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 #E1708 and NEB #L4401 at each template input. No amplification was observed in any of the no template control reactions.




Figure 3: LyoPrime WarmStart Fluorescent LAMP/RT-LAMP Mix (with UDG) enables strong detection of various infectious disease targets

LAMP Infectious Disease

RT-LAMP reactions (25 μl) were performed at 65°C for 30 minutes using 1X primers targeting the indicated viral RNA gene at 5000, 500, or 50 copies per reaction or no template (NTC) as indicated. Fluorescence was monitored every 15 seconds in the SYBR/FAM channel of a real-time thermocycler (Bio-Rad® CFX96). A) Performance was evaluated in quadruplicate for 5000 and 500 copies or twelve replicates for 50 copies of ATCC® synthetic viral RNA (Chikungunya Virus: VR-3246SD; Dengue-2 Virus: VR-3229SD; Zika Virus: VR-3252SD). B) Performance was evaluated in quadruplicate for 5000 and 500 copies or thirty-six replicates for 50 copies of synthetic SARS-CoV-2 RNA. A summary of results from analogous testing conducted by 3 users on SARS-CoV-2 RNA is shown in the table. Each dot represents the time at which the fluorescence signal for a single reaction crosses the instrument-defined threshold. All replicates were detected at each template input for these evaluations unless otherwise indicated (note that dots frequently overlap given similar detection time for the replicates). No amplification was observed in any of the no template control reactions. The LyoPrime WarmStart Fluorescent LAMP/RT-LAMP Mix (with UDG) exhibited robust detection for each infectious disease target.


This product is related to the following categories:
Isothermal Amplification & Strand Displacement Products
This product can be used in the following applications:
Isothermal Amplification

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • L4401S     15 to 25    
      LyoPrime WarmStart™ Fluorescent LAMP/RT-LAMP Mix (with UDG) L4401SVIAL 15 to 25 2 x 50 reactions Not Applicable
  • L4401P     15 to 25    
      LyoPrime WarmStart™ Fluorescent LAMP/RT-LAMP Mix (with UDG) L4401PLATE 15 to 25 1 x 96 reactions Not Applicable

Properties & Usage

Protocols, Manuals & Usage

Protocols

  1. Protocol for LyoPrime WarmStart™ Fluorescent LAMP/RT-LAMP Mix (with UDG) (NEB #L4401)
  2. Protocol for LyoPrime WarmStart™ Fluorescent LAMP/RT-LAMP (with UDG), 96-well plate (NEB #L4401P)

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the optimal LAMP/RT-LAMP amplification temperature?
  2. What is the Mg++ concentration in the WarmStart LAMP/RT-LAMP Master Mix?
  3. What types of input samples/materials are compatible with the LAMP/RT-LAMP mixes?
  4. How fast should I expect a result?
  5. How do I confirm a positive result?
  6. Can I set up my LAMP/RT-LAMP reactions at room temperature?
  7. The WarmStart LAMP Master Mix has some precipitation in the tube after thawing, is this normal?
  8. How do I design LAMP Primers?
  9. Does the WarmStart LAMP/RT-LAMP 2X Master Mix (with UDG) enable carryover prevention/contamination reduction?
  10. Amplification occurred in my NTC sample(s) following isothermal incubation. What happened?
  11. Is a separate incubation step required for RT-LAMP?
  12. What type of purification is recommended for LAMP primers?
  13. What is LAMP and RT-LAMP?
  14. What is the concentration of the LAMP Fluorescent Dye in the LyoPrime WarmStart Fluorescent LAMP/RT-LAMP Mix (with UDG)?
  15. How should the LyoPrime LAMP mix be stored?
  16. How do I rehydrate the LyoPrime LAMP mix? Can it be resuspended at a higher concentration?
  17. When I dissolve the lyophilized product, the solution at first appears turbid. Is this normal?
  18. For L4401P - Do I need to use the entire LAMP plate in a single experiment? How are the 8-well strip tubes separated?
  19. For L4401P - How do I store unused 8-well strip tubes after opening the foil pouch? How long can I keep unused 8-well strip tubes?
  20. For L4401P - How long can I keep the plate or the 8-well strip tubes out of the foil pouch before setting up the reaction?
  21. For L4401P - What is the purpose of yellow shell adaptor?
  22. For L4401P - Can I reuse the yellow shell adaptor?
  23. For L4401P - Why is there a cardboard tray in the package?

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

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

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