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A microsatellite marker parentage identification primer, method and application suitable for Nile tilapia, Olia tilapia and their hybrids

A technology of microsatellite markers and Nile tilapia, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve problems such as no microsatellite paternity identification methods, and achieve High polymorphism, improved efficiency, and improved accuracy

Active Publication Date: 2021-03-16
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Microsatellite paternity testing methods have been reported in aquatic animals such as grass carp, Litopenaeus vannamei, Jian carp, golden squid, mandarin fish, turbot and flounder, although microsatellite paternity has been reported in Nile tilapia. Satellite paternity testing methods were reported, but microsatellite paternity testing methods were not reported in Oria tilapia and Oni tilapia

Method used

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  • A microsatellite marker parentage identification primer, method and application suitable for Nile tilapia, Olia tilapia and their hybrids
  • A microsatellite marker parentage identification primer, method and application suitable for Nile tilapia, Olia tilapia and their hybrids
  • A microsatellite marker parentage identification primer, method and application suitable for Nile tilapia, Olia tilapia and their hybrids

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Experimental program
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Effect test

Embodiment 1

[0031] A kind of microsatellite marker parentage identification method applicable to Nile tilapia, Olia tilapia and hybrids thereof, comprising the steps:

[0032] 1. Family construction: Using the parents of Nile tilapia and Olia tilapia preserved in this laboratory, after they are sexually mature, use artificial insemination to construct 3 Nile tilapia full-sib families, 2 A full-sib family of Olia tilapia and three full-sib families of Olia tilapia (Nile tilapia♀×Olia tilapia♂). Each family was numbered and reared independently, and 2 months after the fry hatched, the tail fin samples of the parents of each family and 32 to 35 offspring were collected as materials for paternity identification.

[0033] 2. Genomic DNA extraction: The caudal fin genomic DNA of the parent and offspring tilapias of each family in step 1 was extracted using an animal tissue micro DNA extraction kit (Magen).

[0034] 3. Screening of polymorphic microsatellite primers: 2 to 3 microsatellite loci ...

Embodiment 2

[0052] Practical application effect evaluation: paternity identification of tilapia "Yuemin No. 1" family

[0053] 1. Family construction and sampling: 5 female Nile tilapias were selected as female parents, and 5 super-male Nile tilapias were selected as male parents and Olia tilapia as female parents. This, the hybrid offspring obtained after backcrossing with the super-male Nile tilapia) was used as the male parent, and 5 full-sib families were constructed by one-to-one pairing, and the parents and 35 offspring of each family were collected after natural seedling production tail fin samples.

[0054] 2. Using the method of step 2 in Example 1 to extract the genomic DNA of the parents and offspring of the full-sib family of the above-mentioned 5 tilapia "Yuemin No. 1"; adopting the method of step 4 in Example 1 to adopt 13 pairs of fluorescently labeled microsatellites The primers were used for PCR amplification and ABI 3730xl sequencer for multiple capillary electrophoresi...

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Abstract

The invention discloses a microsatellite marker paternity identification primer and method suitable for nile tilapia, blue tilapia and a hybrid thereof, and application thereof, and belongs to the technical field of fish breeding. The method screens 13 pairs of fluorescent marker microsatellite primers for paternity identification. The identification method comprises the following steps of: constructing a complete sibling family of the tilapia, the blue tilapia and the hybrid thereof; extracting genomic DNA of parents and progeny of each family; screening polymorphic microsatellite markers; performing PCR amplification on fluorescent marker microsatellite primers; and performing multiple capillary electrophoresis typing and paternity identification. The invention establishes a microsatellite marker paternity identification method suitable for the tilapia, the blue tilapia and the hybrid thereof for the first time, the identification accuracy rate is 100%, and the identification accuracy rate can reach 99.43% in practical application. The method has high identification accuracy and simple and convenient operation, and can be used for guiding population inheritance and family pedigree management of the three tilapia in the breeding process.

Description

technical field [0001] The invention belongs to the molecular marker technology in the technical field of fish breeding, and specifically relates to a microsatellite marker parentage identification primer, method and application suitable for Nile tilapia, Olia tilapia and hybrids thereof. Background technique [0002] Tilapia (Tilapia) is a tropical fish native to Africa, belonging to Perciformes, Perciformes, Cichlidae, Tilapia (also known as Cichlidae, Cichlids genus), about 100 species. Because of its fast growth, miscellaneous food habits, strong fecundity and adaptability, no intermuscular thorns and delicious taste, it became an excellent breeding variety recommended by the Food and Agriculture Organization of the United Nations (FAO) to the world in 1976. At present, tilapia has been cultured in 75 countries and regions, and it has become a worldwide freshwater cultured fish. Since the introduction of Mozambique tilapia (Oreochromis mossambicus) in 1957, my country'...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/16C12Q2531/113C12Q2563/107C12Q2537/143C12Q2565/125
Inventor 刘志刚卢迈新曹建萌高风英可小丽衣萌萌王淼
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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