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Microsatellite family identifying method for diploid misgurnus anguillicaudatus and application of method

An identification method and a diploid technology, which are used in the determination/inspection of microorganisms, biochemical equipment and methods, and resistance to vector-borne diseases, etc., can solve the problems such as microsatellite markers that have not yet been seen, and achieve stable and reliable PCR products Accelerates the acquisition of a large number of effects

Inactive Publication Date: 2014-08-06
HUAZHONG AGRI UNIV
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Problems solved by technology

At present, there has not been any report on the application of microsatellite markers in loach family identification research

Method used

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  • Microsatellite family identifying method for diploid misgurnus anguillicaudatus and application of method
  • Microsatellite family identifying method for diploid misgurnus anguillicaudatus and application of method
  • Microsatellite family identifying method for diploid misgurnus anguillicaudatus and application of method

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Embodiment 1

[0026] 1) Loach ploidy detection and family establishment

[0027] Wild loach parents were collected from three provinces of Hunan, Hubei and Henan, ploidy was determined by flow cytometry, and 20 diploid loach males and females were selected for artificial induced breeding. The ratio of male to female was 1:1, and 20 full-sib families were established. . The fin ray tissues of the parents of each family were clipped and placed in absolute ethanol, and the family information was recorded and stored at -20°C for later use. 20 families were placed in 20 plastic water tanks for separate breeding. After the fry hatched for 20 days, 25-30 fish were randomly selected from each family, fixed with absolute ethanol, and used as samples for family identification.

[0028] 2) Genomic DNA extraction of loach parents and offspring;

[0029] Put the fin ray tissue of each parent and the whole fish (head and viscera) of each offspring into a 2ml centrifuge tube, cut the tissue into pieces ...

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Abstract

The invention discloses a microsatellite family identifying method for diploid misgurnus anguillicaudatus. The method comprises the following steps: acquiring a full-sib family of diploid misgurnus anguillicaudatus; extracting parent and descendant genomic DNA of the diploid misgurnus anguillicaudatus; screening of polymorphism microsatellite marks and synthesizing a primer; conducting PCR amplification on the parent and descendant DNA samples adopting fluorescent PCR reaction; analyzing the parent genetype and the descendant genetype. The method is multiple in selected microsatellite locus allele number, high in polymorphism, and stable and reliable in PCR products; the probability of helping a descendant individual to find the natural male parent and the natural female parent utilizing the microsatellite locus is 97.8% and 99.6% respectively; the integral identification success rate is as high as 97.4%; requirements of family tree analysis and family management in heredity and breeding of the diploid misgurnus anguillicaudatus can be met fully; powerful technical guarantee is provided for molecular breeding research of the species.

Description

technical field [0001] The invention relates to a molecular marker auxiliary technology for fish genetic breeding, in particular to a method for identifying diploid loach families by using fluorescent marker microsatellite markers. Background technique [0002] Loach (Misgurnus anguillicaudatus) is a small freshwater economic fish belonging to Cyprinidae, Loach subfamily, Loach family, Loach genus, widely distributed in China, Japan, Korea, Russia and Southeast Asia and other countries and regions, in my country Except for the Qinghai-Tibet Plateau, it is distributed from south to north. Loach meat is delicious, nutritious, and can be used as medicine. It is known as "ginseng in water". Not only is the demand in the domestic market strong, but it is also one of the main freshwater fish exported by my country's foreign trade. [0003] Loach has strong adaptability to the environment and can be cultured in ponds, paddy fields, net cages and wooden boxes, and also adopts soill...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156Y02A50/30
Inventor 曹小娟王卫民白晓慧田先畅
Owner HUAZHONG AGRI UNIV
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