Pelodiscus sinensis DNA fingerprint identification method

Abstract

The invention relates to a Pelodiscus sinensis DNA fingerprint identification method adopting the technical scheme that high-purity genome DNA is quickly extracted from Pelodiscus sinensis muscle, and then a random amplified primer and an amplified reaction procedure are screened and built. The method comprises the following specific steps: (1) muscle pretreatment comprising drying temperature, moisture content and the like; (2) genome DNA extraction condition optimization, such as water bath time and temperature, centrifugal speed, time and the like; (3) a PCR amplified reaction system and a program establishment comprising a template, a primer and enzyme addition and the like, annealing temperature, time and cycle time design and the like; (4) under the optimal amplified condition, screening different random primers and building the molecular markers of Pelodiscus sinensis in different areas. Compared with the common shape identification technology, the technology is quick and accurate, is not affected by external feeding environmental condition, can quickly identify the Pelodiscus sinensis parent with different regional characteristics and lays the foundation for successively carrying out hybridization breeding.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for identifying Chinese soft-shelled turtle DNA fingerprints Background technique [0002] my country's soft-shelled turtle breeding is developing very rapidly. Its farming scale and technology integration have become the leading industry in freshwater farming. According to incomplete statistics, the annual output exceeds 150,000 tons, with an output value of more than 6 billion yuan. Only Zhejiang Province has an annual demand of 500-100 million yuan. Only the seedlings, and the Jiangnan flowers from the local Taihu product department, the annual supply capacity of the seedlings is only 10 million, a large number of seedlings or eggs are entered from Taiwan, Thailand and other places, resulting in high-quality Chinese seedlings resources resources Affected by the threat of serious mixing and decline, germplasm degeneration appeared, which brought great harm to the soft-shelle...

AI Technical Summary

Problems solved by technology

[0002] my country's soft-shelled turtle breeding is developing very rapidly. Its farming scale and technology integration have become the leading industry in freshwater farming. According to incomplete statistics, the annual output exceeds 150,000 tons, with an output value of more than 6 billion yuan. Only Zhejiang Province has an annual demand of 500-100 million yuan. Only the seedlings, and the Jiangnan flowers from the local Taihu product department, the annual supply capacity of the seedlings is only 10 million, a large number of seedlings or eggs are entered from Taiwan, Thailand and other places, resulting in high-quality Chinese seedlings resources resources Suffered from severe mixing and recession threats, and signs of germplasm degradation appeared, which brought great harm to the soft-shelled turtle industry

On the one hand, due to the neglect of source work such as germplasm protection and purification and breeding, domestic soft-shelled turtle populations and strains are mixed, inbreeding is serious, the genetic diversity of offspring is continuously reduced, harmful genes are continuously purified, and the germplasm of soft-shelled turtles is seriously degraded. Severely degraded good commodity traits, reduced disease resistance and immunity, slow growth, reduced quality of commercial soft-shelled turtles, early sexual maturity, and smaller size of sexually mature individuals; decreased quality of commercial soft-shelled turtles, decreased meat quality and umami taste, and decreased acceptance by the common people , there is a phenomenon of "high yield but not good harvest"; on the other hand, the quality of seedlings imported from Taiwan, Thailand and other places is uneven, which brings the threat of mixed degeneration of soft-shelled turtle germplasm resources in this region

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