Specific primers of acanthopagrus latus microsatellite marker and application of specific primer

A technology of microsatellite marking and yellowfin roughy, which is applied in the field of microsatellite marking, can solve the problems such as unfavorable breeding process of yellowfin roughy species, and achieve convenient breeding of improved species and family management, high polymorphism, and economical The effect of experiment cost

Active Publication Date: 2018-12-21
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

At present, some scholars have developed a large number of SSR markers of yellowfin roughy, but there are few reports on the applica...

Method used

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  • Specific primers of acanthopagrus latus microsatellite marker and application of specific primer
  • Specific primers of acanthopagrus latus microsatellite marker and application of specific primer
  • Specific primers of acanthopagrus latus microsatellite marker and application of specific primer

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1 Screening and synthesis of specific primers for yellowfin roughy microsatellite markers

[0046] Select sequences containing microsatellite markers from the yellowfin seabream transcriptome library to design primers, screen the unigene containing yellowfin seabream microsatellite repeat sequences through transcriptome sequencing, and then detect them from the yellowfin seabream transcriptome library Design microsatellite-specific primers to the sequences containing microsatellite sites, and detect their polymorphisms. After screening, 8 pairs of primers with clear bands and high polymorphisms were finally selected as primers for family identification. Synthesized by Bioengineering Co., Ltd., the 5' ends of the forward primers of each pair of microsatellite primers were modified with three different fluorescent groups: FAM, HEX, and TAMRA (see Table 1). The primers were purchased from Shanghai Sangon Bioengineering Co., Ltd. Ltd Synthetics.

[0047] Table 1 In...

Embodiment 2

[0049] Example 2 Yellowfin roughy microsatellite paternity identification method

[0050] (1) Establishment of relationship between parents and offspring of yellowfin roughy seabream

[0051] In this experiment, 112 yellowfin roughy broodstock were sourced from wild seedlings caught in the sea in Yangjiang City, Guangdong Province, and bred in the experimental base of Hainan Tropical Fisheries Research and Development Center, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences. The 393 offspring were derived from the mixed family of yellowfin roughy obtained through artificial insemination in December 2017. Fin rays and whole fish were collected from parents and offspring, respectively, and stored in absolute ethanol for later use.

[0052] (2) Extraction of genomic DNA from parents and offspring of yellowfin roughy seabream

[0053] Using the Magen Animal Tissue DNA Extraction Kit, process 20-50mg tissue samples into as small fragments as possi...

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Abstract

The invention discloses specific primers of an acanthopagrus latus microsatellite marker. The specific primers include 8 pairs of primers: AL15, AL20, AL51, AL01, AL37, AL18, AL14 and AL49; the primers have high polymorphism, and the PCR product is stable and reliable. The invention also discloses a microsatellite paternity testing method of acanthopagrus latus, wherein a paternity testing molecular method of acanthopagrus latus is established for the first time, and the method has the advantages of accurate test and low price; the invention further discloses application of the specific primers of the acanthopagrus latus microsatellite marker in the paternity test of acanthopagrus latus.

Description

technical field [0001] The invention belongs to the technical field of microsatellite markers, and in particular relates to a specific primer and application of microsatellite markers for yellowfin roughy sea bream. Background technique [0002] Yellowfin seabream (Acanthopagrus latus) belongs to fish class Pisces, perciformes Perciformes, sea bream family Sparidae, sea bream genus Acanthopagrus, and is widely distributed in the Red Sea, Arabian Sea, Indian Ocean, western Pacific coast and China Taiwan, Fujian, Guangdong, Guangxi coastal. [0003] Yellowfin roughy lives in coastal waters and estuaries. It is omnivorous and is an important economic fish. The breeding cycle of yellowfin roughy is relatively long, and it takes one and a half to two years to reach the market size of about 5 taels, which greatly limits fishermen's enthusiasm for breeding this species. In recent years, due to environmental pollution, overfishing, etc., the natural population of yellowfin roughy ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 朱克诚张殿昌江世贵刘宝锁郭华阳郭梁张楠
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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