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Microsatellite marking method for parentage identification of coreius guichenoti

A technology of microsatellite marking and copperfish, which is applied to biochemical equipment and methods, and microbial measurement/inspection, can solve the problems of no application of microsatellite marking, optimize the setting of annealing temperature, reduce time cost, The effect of improving the amplification effect

Active Publication Date: 2018-12-14
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

But at present, there is no report on the application of microsatellite markers in the paternity test of copperfish

Method used

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  • Microsatellite marking method for parentage identification of coreius guichenoti
  • Microsatellite marking method for parentage identification of coreius guichenoti
  • Microsatellite marking method for parentage identification of coreius guichenoti

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Embodiment 1

[0023] Below in conjunction with example, the present invention will be further described.

[0024] A microsatellite marker method for paternity identification of copperfish, the steps of which are:

[0025](1) Extract the DNA of the parent and offspring individuals of copperfish: collect 8 family samples (including 22 true parents and 297 offspring), and select 20 non-parental samples as candidate parents, of which 8 family samples Including 5 full-sib families, 1 half-sib family and 2 mixed families. Use absolute ethanol to preserve the fin ray tissue of the parent copperfish and the whole fish individual samples of the offspring that have just hatched out of the membrane, and use the Tissue DNA Kit (D3396) kit from Omega Bio-Tek Company to extract the genomic DNA of the sample. The specific steps are as follows : Select about 30mg of round mouth copper fin ray or whole fish sample, wash with double distilled water, put it into a 1.5mL sterilized centrifuge tube, cut it int...

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Abstract

The invention discloses a microsatellite marking method for parentage identification of coreius guichenoti. The microsatellite marking method comprises the steps: (1) extraction of a sample DNA from acoreius guichenoti family; (2) screening of coreius guichenoti polymorphic microsatellite primers: according to the amplification effect of wild individuals of coreius guichenoti, screening to obtain20 pairs of polymorphic microsatellite primers from 40 pairs of primers recorded in literature; (3) coreius guichenoti fluorescence-marked microsatellite PCR amplification and multiple capillary electrophoresis: marking 20 pairs of screened microsatellite primers marked with different fluorescent substances, carrying out landing PCR amplification of the DNA sample of the coreius guichenoti familyby the fluorescent primers, and carrying out capillary electrophoresis of multiplex combinations formed by the PCR amplified products; and (4) parentage identification and analysis of coreius guichenoti: according to the electrophoretic results, reading genotype data of parents and offspring, and determining the parent-offspring relationship between the to-be-tested offspring and the parents. Themethod is simple to implement and easy, and provides a new technical means for coreius guichenoti family management, population genetic management and evaluation of reproductive and releasing effects.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and more specifically relates to a microsatellite marker method for paternity identification of copperfish. Background technique [0002] Roundmouth copperfish (Coreius guichenoti (Sauvage et Dabry)), belonging to Cypriniformes (Cypriniformes), Cyprinidae (Cyprinidae), Gobioninae (Gobioninae), copper fish (Coreius), distributed in the upper reaches of the Yangtze River and the elegant In large tributaries such as the Longjiang River and the Wujiang River, there are typical river migratory fish and fish that lay drifting eggs. Roundmouth copperfish is a unique fish in the upper reaches of the Yangtze River, and it is also an important economic fish. The spawning grounds of copperfish are only found in the middle and lower reaches of the Jinsha River and the lower reaches of the main stream of the Yalong River. With the gradual implementation of hydropower development in the upper reache...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2525/151C12Q2565/125C12Q2563/107C12Q2537/143
Inventor 何勇凤杨德国朱永久吴兴兵
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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