PCR primer for identifying species specificity of microhyla mixtura

Abstract

The invention discloses a PCR primer for identifying the species specificity of microhyla mixtura. A COI gene target fragment comprises an upstream primer sequence L1 5'-GGC CCC CGC TAT ACG AAA AAC-3', and a downstream primer sequence L1 5'-GGC AGG GGT GAA GTT GTCTGG G-3'. DNA of the microhyla mixtura is extracted from the muscular tissue of the microhyla mixtura, the extracted DNA is subjected toPCR amplification and then is subjected to agarose gel electrophoresis detection, and then whether the microhyla mixtura is true is identified according to the fact whether strips occur in an agarosegel electrophoresis detection result. Compared with traditional DNA sequencing and sequence alignment, the identifying mode has the advantages that the identifying mode is simple and accurate, whether the microhyla mixtura is true can be identified through one PCR amplification and agarose gel electrophoresis detection, the detection time is reduced, the detection efficiency is improved, and theidentifying mode has the effect of convenience and economical efficiency.

Description

technical field [0001] The invention relates to the field of biological detection and identification, in particular to a specific PCR primer for identifying the species-specificity of A. Background technique [0002] There are many kinds of amphibians in the genus Aphidia, including Aphidia japonica, Adiaba sylvestris, Amosia spp., Amosia chinensis, and Amosia japonica, etc. They have a wide range of distribution, and their distribution areas often overlap. [0003] The scientific name of Hezhengji frog Microhyla mixtura , is an endemic species in China, and is an amphibian of the genus Rana in the family Aphididae. It is distributed in Anhui, Hubei, Sichuan, Shaanxi, Guizhou and other places, and mostly lives in small puddles in mountainous areas, alpine rice fields and nearby. Its survival altitude ranges from 140 to 1700 meters. In the process of collecting the specimens of Syndromic Frogs, the researchers of our research group found that there are great differences in...

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Problems solved by technology

In the process of collecting the specimens of the Syndromic Frog, the researchers of our research group found that the morphology and size of the Syndromic Frog in different collection locations are quite different, and the differentiation of species or subspecies may have occurred, so the identification is based solely on the shape and size characteristics. It is quite difficult to find the Frogs of Syndrome, and in some collection sites, it is found that Frogs of Syndromes live together with Frogs of Ornaments and Frogs of Flowers. Syndromic Rana, so we need to design a specific primer at the molecular level to identify the species specificity of Syndromic Rana

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