Loop-mediated isothermal amplification (LAMP): Facilitates running amplification reactions in difficult test environments, enabling use outside of a high-tech laboratory.
Master Mix Format: Simplifies reaction setup and minimizes optimization required to produce the best assay results.
High Thermal Stability: Highly thermostable enzyme (up to 90°C for 5 min) enables preheating of reactions containing purified target DNA which, depending on the target, may increase assay sensitivity and decrease time to results.
Elevated LAMP Reaction Temperature (68-74°C): Improves primer specificity and reduces background amplification depending on the DNA target.
Freeze-dry Compatible: Enables generation of room temperature stable test kits through lyophilization.
Customizable: Once primers are optimized with the LavaLAMP™ DNA Master Mix, we can work with you to generate custom master mixes further optimized for your target and/or test needs – contact us
The LavaLAMP™ DNA Master Mix is intended to simplify development and optimization of DNA LAMP (loop-mediated isothermal amplification) reactions. LAMP kits are commonly available as multi-component kits that require optimization (e.g. MgSO4, betaine, enzyme as well as temperature, primer concentration, etc.). The LavaLAMP™ DNA Master Mix greatly simplifies reaction optimization by limiting optimization to target specific components/conditions such as LAMP primer design, target concentration and reaction temperature. Additionally, this master mix is heat stable at 90°C for ≤5 minutes, which enables the addition of a reaction preheating step which may, depending on the target, increase assay sensitivity and decrease time to results. Need more information? See our FAQs.
Figure 1. Simplified overview of the loop-mediated isothermal amplification (LAMP) process. For simplicity, 4/6 potential LAMP primers are illustrated. The inclusion of two additional primers, F-Loop; Forward Loop Primer and B-Loop; Backward Loop Primer, often significantly enhances amplification. For more information, please visit the Eiken website.
Figure 2. Illustration of the differences in LAMP reaction setup and potential optimization parameters when using a master mix vs. a component kit format. Panel A illustrates the components added to a LAMP reaction (left) when using the LavaLAMP DNA Master Mix and the various parameters than can be optimized (right). Panel B also illlustrates reaction setup (left) and potential optimization parameters (right) when using a standard LAMP component kit.
Figure 3. Loop-mediated isothermal amplification (LAMP) with real-time fluorescent detection of amplified products. LAMP reactions were set up using the indicated kits according to manufacturer’s recommendations. Target DNA (C. difficile) at varying input amounts, tcdA target LAMP primers, and Green Fluorescent Dye (LavaLAMP Kit) were included in all reactions. Reactions were run on a CFX96 Thermal Cycler (Bio-Rad) at the following temperatures: LavaLAMP; 68°C; other kits at the recommended 65°C and fluorescence was measured over 60 minutes to determine the TTR. NTC = No Target Control.
Figure 4. Testing the thermostability of the LavaLAMP™ DNA Master Mix. Two sets of triplicate LAMP reactions were set up using the indicated amounts of target DNA (C. difficile), tcdA target LAMP primers, and Green Fluorescent Dye. One set of reactions was preheated to 90°C for 5 minutes, and then all reactions were incubated for 60 minutes on a CFX96 Thermal Cycler (Bio-Rad) at 68°C and fluorescence was measured during the course of the reactions to determine the TTR. NTC = No Target Control.
Figure 5. Testing the performance of lyophilized LavaLAMP™ DNA Master Mix. The LavaLAMP DNA Master Mix (12.5 µL) was aliquoted into tubes and lyophilized using a VirTis Wizard 2.0. After one day, each lyophilized tube was reconstituted in 12.5 µL water. Then six replicate LAMP reactions per condition were set up using the reconstituted lyophilized master mix and the standard LavaLAMP DNA Master Mix (Controls). The indicated amounts of target DNA (S. aureus), clfA target LAMP primers, and Green Fluorescent Dye were added to the reactions. All reactions were incubated for 60 minutes in a CFX96 Thermal Cycler (Bio-Rad) at 68°C and fluorescence was measured during the reactions to determine the TTR. NTC = No Target Control.
Figure 6. Analysis of day to day variability. On three different days, six replicate LAMP reactions were set up per condition using the indicated amounts of target DNA (M13mp18), M13mp18 target LAMP primers, and Green Fluorescent Dye. On each day the reactions were run on a CFX96 Thermal Cycler (Bio-Rad), amplification was monitored in real-time, and then results were analyzed to determine the TTR. NTC = No Target Control.