Gene mutation detection method using connection-based loop-mediated isothermal amplification technology

Abstract

The invention provides a gene mutation detection method using a connection-based loop-mediated isothermal amplification technology. The detection method comprises the following steps: (1) designing two DNA hairpin probes H1 and H2 by taking target DNA as a template; (2) in the presence of target, performing annealing hybridization of the target DNA with the H1 and H2; and in the case of complete complementation of the 3' end of H1, connecting the H1 and H2 by ligase to form a double-dumbbell DNA structure; (3) performing a loop-mediated isothermal amplification reaction by taking the double-dumbbell DNA as a template to obtain a loop-mediated isothermal amplification product; (4) adding DNA double-chain specific dye SYBR GreenI into the loop-mediated isothermal amplification product to enhance the fluorescence signal, and implementing quantitative detection of gene mutation through fluorescence signal detection. The gene mutation detection method provided by the invention has remarkably low lower detection limit and relatively high selectivity and specificity.

Description

technical field [0001] The invention belongs to the fields of molecular biology and nucleic acid chemistry, and in particular relates to a gene mutation detection method based on a connection-based loop-mediated isothermal amplification technology. Background technique [0002] From the molecular level, gene mutation refers to the change of base pair composition or sequence in gene structure. Gene mutations can occur at any stage of development, usually during the DNA replication period, that is, cell division interphase, including mitotic interphase and meiotic interphase. Gene mutation is also one of the important factors in biological evolution, so the study of gene mutation is not only In addition to its theoretical significance, it also has extensive biological significance. Gene mutation provides mutant types for genetic research and materials for breeding work, so it also has practical significance in scientific research and production. With the in-depth study of mol...

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Problems solved by technology

However, there is no single-base mutation detection using improved LAMP

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