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What is Loop-mediated Isothermal Amplification using Loop Primers?

Learn how pLAMP simplifies primer design, enhances specificity, and improves amplification homogeneity, making it ideal for medical diagnostics, pathogen detection, and genetic research

April 1, 2024

Loop-mediated isothermal amplification (LAMP) has emerged as a powerful and versatile nucleic acid amplification technique, finding extensive applications in medical diagnostics, pathogen detection, food safety monitoring, and environmental surveillance. In comparison to PCR (Polymerase Chain Reaction), LAMP offers numerous advantages, including simplicity, rapidity, minimal equipment requirements, and robustness against amplification inhibitors.

Typically, LAMP involves the utilization of 4-6 primers, comprising two sets of specific primers: outer primers (F3 and B3) and inner primers (FIP and BIP), along with 1-2 loop primers. The outer primers bind to regions flanking the target sequence, while the inner primers bind within the target sequence. Loop primers, on the other hand, target loop structures formed during the LAMP reaction. The design of these primers is critical for efficient amplification, ensuring specificity and compatibility with the isothermal conditions required for LAMP.

The role of loop primers in LAMP is to enhance the speed and efficiency of amplification. As the LAMP reaction commences, loop primers bind to the already-formed LAMP amplification products, facilitating the rapid progression of the amplification process. Through this mechanism, loop primers accelerate the attainment of equilibrium in the reaction, thereby increasing the speed and yield of LAMP amplification.

SBS Genetech's unique Premium LAMP (pLAMP), based on the distinctive Bst P DNA/RNA Polymerase, represents a significant advancement in LAMP technology by integrating Helicase enzyme into the reaction mixture. This addition offers several advantages over traditional LAMP techniques. Notably, pLAMP can operate without the need for outer primers (F3/B3) due to the presence of Helicase enzyme, which facilitates the unwinding of DNA strands. Consequently, pLAMP reactions can be initiated solely with inner primers (FIP/BIP), streamlining primer design and reaction setup.

Furthermore, Helicase enzyme assists in the unwinding of DNA strands during amplification, thereby reducing the concentration of FIP/BIP primers required for efficient amplification. This reduction in primer concentration minimizes non-specific amplification and enhances the uniformity of amplification across target sequences, leading to improved specificity, sensitivity, and reproducibility of pLAMP assays.

In summary, Premium LAMP (pLAMP) harnesses the capabilities of Helicase enzyme to streamline primer design, reduce non-specific amplification, and improve amplification homogeneity. These advancements make pLAMP an attractive option for various applications, including diagnostic testing, pathogen detection, and genetic research, further cementing LAMP's position as a leading molecular diagnostic tool.

For more information on our Premium LAMP (pLAMP) and Bst P DNA/RNA Polymerase, visit the Science Journal article dated April 13, 2023, or contact us to discover how SBS Genetech can accelerate your research journey.

 

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