Microsatellite marker primers for identification of Acipenser dabryanus family and identification method thereof

A technology of microsatellite markers and sturgeon, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Achieve the effect of preventing the decline of population quality, strong gene correlation, and high polymorphism

Inactive Publication Date: 2017-07-11
FISHERIES INST SICHUAN ACADEMY OF AGRI SCI
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Problems solved by technology

However, the method of cultivating a single family alone not only takes up a lot of space, but also has high labor intensity and high cost of breeding equipment. More importantly, different environmental effects will affect the growth traits of different families; and traditional p

Method used

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  • Microsatellite marker primers for identification of Acipenser dabryanus family and identification method thereof
  • Microsatellite marker primers for identification of Acipenser dabryanus family and identification method thereof

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Embodiment 1

[0033] Example 1 Acquisition and verification of microsatellite marker primers for family identification of Dabry's sturgeon:

[0034] Dabry's sturgeon sample collection and RNA extraction:

[0035] Two healthy Dabry's sturgeons were selected from the Sichuan Fisheries Research Institute, and their gonads were dissected for total RNA extraction, followed by sequencing, splicing and assembly of the Dabry's transcriptome and searching for microsatellite sites. After screening, 18 pairs of primers with clear bands and high polymorphism were finally selected as primers for family identification. The primers were synthesized in Shanghai Sangon Bioengineering Co., Ltd.

[0036] Table 1 Information of microsatellite primers used for family identification of Acipenser dabryi

[0037]

[0038]

Embodiment 2

[0039] The identification method of embodiment 2 Dabry's sturgeon microsatellite families

[0040] (1) DNA extraction from the parent and offspring of Dabry's sturgeon

[0041] (1) Establishment of Dabry's sturgeon family

[0042] Selected 43 sexually mature Dabry's sturgeons from the Sichuan Fisheries Research Institute as the candidate parents for this identification, and selected 43 F2 generation bred in 2014 and 24 F2 generation bred in 2015, a total of 67 juveniles were selected as the identification parents. Offspring. At the same time, in order to ensure the reliability of the identification method, 20 Dabry's sturgeon offspring (from two full-sibling breeding families, each with 10 fry in each family) were selected as positive references. to assess the accuracy of the identification method.

[0043] (2) Extraction of Genomic DNA from Parents and Progeny of Dabry's Acipenser

[0044]The preparation of Dabry's sturgeon DNA was carried out by CTAB method, and the spec...

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Abstract

The invention relates to microsatellite marker primers for identification of an Acipenser dabryanus family. Through a transcriptome sequencing technology, Unigenes of the screened Acipenser dabryanus are screened, microsatellite loci in the Unigenes sequence are detected, according to flanking sequences at two ends of each one of the microsatellite molecular marker loci, specific primers of the molecular marker are labeled, the specific primers are subjected to PCR amplification, the stability and polymorphism of the amplification result are detected, and 18 effective microsatellite markers are acquired. Through the microsatellite marker primers, according to the polymorphism of the microsatellite marker primers, a parentage identification platform of Acipenser dabryanus is built and the basis is provided for Acipenser dabryanus selection and breeding matching.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to a microsatellite marker primer and an identification method for the family identification of Dabry's sturgeon. Background technique [0002] Dabry's sturgeon, also known as Changjiang sturgeon and Sarba, is a unique species in my country. It is mainly distributed in the main stream of the Yangtze River and the lower reaches of the Jinsha River, and the Yibin-Hejiang section of Sichuan has a higher yield. In the past 20 years, due to the construction of Gezhouba Project, water pollution, overfishing and other reasons, the wild resources of Dabry's sturgeon have been extremely scarce. In 1988, it was listed as a national first-level key protected animal and a first-level urgently protected fish in the upper reaches of the Yangtze River. Domestication and breeding of wild resources of Dabry's sturgeon, and artificial reproduction after reaching sexual maturity are importan...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6869C12Q1/6888C12Q2600/156C12Q2525/151C12Q2563/107C12Q2535/122
Inventor 陈叶雨刘亚杜军龚全赵刚林珏赖见生
Owner FISHERIES INST SICHUAN ACADEMY OF AGRI SCI
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