Procypris rabaudi Tchang test kit and microsatellite PCR identification method thereof

A technology of paternity identification and identification method, which is applied in the field of Iwahara carp paternity identification kit and its microsatellite PCR identification, can solve the problems that microsatellite markers have not been found, and achieve a stable amplification effect

Active Publication Date: 2020-12-25
WATER ENG ECOLOGICAL INST CHINESE ACAD OF SCI
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Problems solved by technology

At present, there is no research report on the application of microsatellite markers in paternity testing of Iwahara carp

Method used

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  • Procypris rabaudi Tchang test kit and microsatellite PCR identification method thereof
  • Procypris rabaudi Tchang test kit and microsatellite PCR identification method thereof
  • Procypris rabaudi Tchang test kit and microsatellite PCR identification method thereof

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Embodiment

[0032] Below in conjunction with example, the present invention will be further described.

[0033] Iwahara carp paternity identification kit and microsatellite PCR identification method thereof, the steps are:

[0034] (1) Genomic DNA extraction of individual samples of Iwahara carp: samples of 4 parents and 86 offspring fry of the family obtained through artificial pairing were taken out, and genomic DNA was extracted according to the high-salt method. The specific steps are as follows: cut about 0.5g of fin ray tissue (the offspring fry individuals use all the fry), wash the alcohol attached to the preserved sample with double distilled water, fully cut it into pieces, and put it into a 1.5mL centrifuge tube; Add 500 μL of cell lysate and 6 μL of proteinase K to the centrifuge tube, digest overnight in a 55°C water bath, and shake several times in the first 30 minutes; add 500 μL of sodium chloride (4.5mol / L), 300 μL of chloroform, and shake at medium speed Homogenize for ...

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Abstract

The invention discloses a procypris rabaudi Tchang test kit and a microsatellite PCR identification method thereof. The method comprises the steps of extracting genome DNA of a procypris rabaudi Tchang individual sample; performing PCR amplification; marking the 5'end of a forward primer of each pair of 20 microsatellite primers in a procypris rabaudi Tchang paternity test primer group with a fluorescent substance, performing gradient PCR amplification on the obtained DNA genome by using each marked fluorescent primer in the primer group, performing capillary electrophoresis on an amplification product on a sequencer, reading the allele size of each individual, and obtaining genotyping data; and performing paternity test analysis of the procypris rabaudi Tchang family: performing parent and filial generation data analysis based on the genotype data, and determining the paternity relationship between the filial generation and the parent according to the correlation between the filial generation genotype and the parent genotype. The method is simple, quick, efficient and economical, and can provide a technical approach for family management, population genetic variation research andproliferation and release effect evaluation of procypris rabaudi Tchang breeding populations.

Description

technical field [0001] The invention relates to the technical field of fish breeding and molecular markers, and relates to a technique for paternity identification of Iwahara carp based on microsatellite markers, in particular to a Iwahara carp paternity identification kit and a microsatellite PCR identification method thereof. Background technique [0002] Procypris rabaudi Tchang belongs to Cypriniformes, Cyprinidae, Cypininae, Procypris, local names rock carp, black carp, rock carp, etc. Distributed in the main stream and tributaries of the Yangtze River above Yichang, it is a bottom fish inhabiting rock crevices, especially in the Jialing River, Minjiang River and Jinsha River. It is now one of the wild and valuable economic fish in Sichuan, Chongqing and other places. Due to the relatively narrow distribution area and small population of Iwahara carp, coupled with overfishing, the pollution of the Yangtze River waters, and the construction of dams and gates in the upper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2563/107C12Q2565/125Y02A40/81
Inventor 阙延福李伟涛朱滨胡兴坤徐念熊美华邵科汪鄂洲田华
Owner WATER ENG ECOLOGICAL INST CHINESE ACAD OF SCI
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