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Spinibarbus sinensis paternity test kit based on microsatellite markers, and method thereof

A microsatellite marker and paternity test technology, which is applied to the Chinese barb paternity test kit and the microsatellite PCR identification field, can solve problems such as microsatellite markers that have not yet been found, and achieve the effect of stable amplification effect

Active Publication Date: 2021-01-15
WATER ENG ECOLOGICAL INST CHINESE ACAD OF SCI
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  • Abstract
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Problems solved by technology

At present, there is no research report on the application of microsatellite markers in paternity identification of Chinese barb

Method used

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  • Spinibarbus sinensis paternity test kit based on microsatellite markers, and method thereof
  • Spinibarbus sinensis paternity test kit based on microsatellite markers, and method thereof
  • Spinibarbus sinensis paternity test kit based on microsatellite markers, and method thereof

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Embodiment

[0027] Below in conjunction with example, the present invention will be further described.

[0028] A method for paternity identification of Chinese spiny barb based on microsatellite markers, the steps of which are:

[0029] (1) Genomic DNA extraction of individual samples of barb barb: The samples of 6 parents and 96 offspring fry of the family obtained through artificial pairing were taken out, and the 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 Homo...

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Abstract

The invention discloses a spinibarbus sinensis paternity test kit based on microsatellite markers, and a method thereof. The method comprises the following steps of: extracting the genome DNA of an individual sample of a spinibarbus sinensis; performing PCR amplification: marking the 5'ends of forward primers of each pair of primers of 15 pairs of microsatellite primers in a primer group for spinibarbus sinensis paternity test with fluorescent substances, performing gradient PCR amplification on the obtained DNA genome by utilizing the marked fluorescent primers in the primer group, performingcapillary electrophoresis of an amplification product on a sequencer, reading the allele size of each individual, and obtaining genotyping data; and performing paternity test analysis of the spinibarbus sinensis family: performing parent and filial generation data analysis based on the genotype data, and determining the parent-child relationship between the filial generation and the parent according to the correlation between the filial generation genotype and the parent genotype. The method disclosed by the invention is simple and rapid, can be popularized and applied in the aspects of germplasm identification and family genetic management of the spinibarbus sinensis, and provides a new technical means for proliferation and release effect evaluation.

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 Chinese spiny barb based on microsatellite markers, specifically a Chinese spiny barb paternity identification kit and a microsatellite PCR identification method thereof. Background technique [0002] Chinese barbed barb (Spinibarbus sinensis) belongs to Cypriniformes (Cypriniformes), Cyprinidae (Cyprinidae), spinibarbus (Spinibarbus), commonly known as "Qingbo", is one of the important economic fish in the upper reaches of the Yangtze River. The proportion of catches was higher in the Wujiang River Basin and the middle and lower reaches of the Chishui River. Chinese barb is big, lively, likes the rapids with clear water, grows fast, and is omnivorous. It is deeply loved by the people. However, in recent years, due to factors such as overfishing, serious water pollution, and habitat destruction caused by the c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/124C12Q2600/156C12Q2525/151C12Q2565/125C12Q2563/107Y02A40/81
Inventor 阙延福李伟涛熊美华徐念汪鄂胡兴坤田华朱滨
Owner WATER ENG ECOLOGICAL INST CHINESE ACAD OF SCI
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