Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification

Loop-mediated isothermal amplification (LAMP) is restricted to detecting a single target, limiting the usefulness of this method. To achieve multiplex LAMP-based detection, we developed a novel approach we called the multiple endonuclease restriction real-time–LAMP assay. In this system, the LAMP forward or backward inner primers contain 5′ end short sequences that are recognized by the restriction endonuclease Nb.BsrDI, and the new forward or backward inner primers were modified at the 5′ end with a fluorophore and in the middle with a dark quencher.

Source: Journal of Molecular Diagnostics - June 19, 2015 Category: Pathology Authors: Yi Wang, Yan Wang, Ruiting Lan, Huaqing Xu, Aijing Ma, Dongxun Li, Hang Dai, Xuejiao Yuan, Jianguo Xu, Changyun Ye Tags: Regular article Source Type: research