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Chinese giant salamander group dividing method based on mitochondrial DNA sequence

A DNA sequence and group division technology, applied in the field of biological identification, can solve the problems of difficult group division standards, no group markers, unstable body surface patterns, etc., and achieve the effects of easy operation, low experimental operation cost, and easy mastery

Inactive Publication Date: 2016-10-26
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI +1
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, some workers identify giant salamanders of different geographical groups by their body surface patterns, but the body surface patterns are not stable and have no fixed signs, so it is difficult to use them as a standard for group division
There are some reports at home and abroad that use mitochondrial DNA to analyze a small number of samples and find that there are certain genetic differences in Chinese giant salamander populations, but no clear population markers have been proposed

Method used

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  • Chinese giant salamander group dividing method based on mitochondrial DNA sequence
  • Chinese giant salamander group dividing method based on mitochondrial DNA sequence
  • Chinese giant salamander group dividing method based on mitochondrial DNA sequence

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Embodiment

[0041] (1) Sample collection and DNA extraction

[0042] Chinese giant salamander individuals were collected from Xing'an in Guangxi (GXXA), Longshan in Hunan (HNLS), Jing'an in Jiangxi (JXJA), Huangshan in Anhui (AHHS), Laifeng in Hubei (HBLF), and Lishui in Zhejiang (ZJLS). The part or tissue collected can be a small amount of fin rays, blood, muscle, fresh molted skin, or oral epithelial cells scraped with a sterilized cotton swab, etc. Genomic DNA was extracted using DNA extraction kit or phenol-chloroform method. Use a spectrophotometer to measure the purity and content of the DNA, and store it at -20°C for future use. The DNA concentration was adjusted to 10-100ng / μL when used.

[0043] (2) Primer synthesis

[0044] The following primers were synthesized at Shanghai Sangon Biotechnology Co., Ltd.:

[0045] L14764: 5′-ATGAAACAGGGTCAAAGCAATCCAAC-3′

[0046] H16062: 5'-AGATGAGGGCAGACTCAGTTATG-3'.

[0047]Dissolve and dilute the primers to a concentration of 10 μmo / L w...

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Abstract

The invention discloses a Chinese giant salamander group dividing method based on a mitochondrial DNA sequence. The method includes: (1), collecting a giant salamander sample, and extracting genome DNA; (2), synthesizing a giant salamander specific mitochondrial DNA primer; (3), performing PCR (polymerase chain reaction) amplification on the giant salamander sample, and sequencing PCR amplification products; (4), comparing an acquired sequence with a reference sequence; (5), according to sequence features of corresponding sites, judging a group to which the sample belongs. The giant salamander mitochondrial DNA sequence is used as a sequence marker for differentiating Chinese giant salamander groups, and target groups can be identified quickly and accurately by comparing the giant salamander mitochondrial DNA sequence with sequence features of the reference sequence. The method can be used for identifying giant salamander individuals in giant salamander farms to determine origin of the individuals; the method can assist in breeding selection during parental breeding. The method can also be applied in identifying multiplication and releasing samples, determining groups to which the samples belong and then releasing and the samples to proper regions, so that germplasm hybridization of wild groups by released groups is avoided.

Description

technical field [0001] The invention belongs to the technical field of biological identification, and in particular relates to a method for grouping Chinese giant salamanders based on mitochondrial DNA sequences. Background technique [0002] The Chinese Giant Salamander (Andrias davidianus) is the largest amphibian among the existing amphibians. The largest individual can reach a length of more than 1 meter and a weight of more than 8 kilograms. The Chinese giant salamander is only distributed in my country and is endemic to my country. The distribution areas include Beijing, Hebei, Henan, Shanxi, Shaanxi, Gansu, Qinghai, Sichuan, Guizhou, Hubei, Hunan, Anhui, Jiangsu, Zhejiang, Jiangxi, Fujian, Guangdong and Guangxi, etc. Province City District. Due to its important scientific research and economic value, the market demand is very large, leading to overfishing of giant salamanders in China, coupled with the reduction of habitats, the current wild resources of giant salaman...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 汪登强梁志强危起伟伍远安
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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