Method applied to identification of pacific oyster family

A Pacific oyster and identification method technology, applied in the field of Pacific oyster family identification, can solve the problems of weak genetic research, influence, lack of improved species, etc., and achieve the effect of high resolution of dissolution curve, high stability and good repeatability

Active Publication Date: 2014-02-26
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

Although my country is a big oyster farming country, it is not yet a strong oyster farming country. There are problems such as relatively lagging behind in genetic improvement rese

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  • Method applied to identification of pacific oyster family
  • Method applied to identification of pacific oyster family
  • Method applied to identification of pacific oyster family

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Embodiment 1

[0021] 1) Primer design and optimization.

[0022] In the SNP database obtained by the Illumina GoldenGate method, search for the first and second SNP mutation sites (C / T&G / A, C / A&G / T), and use Primer3.0 (http: / / primer3 .ut.ee / ) to design primers. When designing primers, try to avoid hairpin structures and primer dimers. The primer length is 18-30bp, the annealing temperature is 55-65°C, and the length of the amplified fragment is 50-180bp. Genomic DNA of 6 individual Pacific oysters was extracted using Qiagen kits, quantitative analysis of DNA was performed with PicoGreen fluorescent dye kits, and the concentration of sample DNA was adjusted to 5 ng / μL for screening and optimization of SNP markers. Using the quantified DNA as a template, use Roche LightCycler480 for HRM detection, by changing the annealing temperature and Mg 2+ The concentration screened primers with high amplification efficiency and clear melting curve, and obtained the optimal PCR reaction system and the...

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Abstract

The invention discloses a method applied to identification of pacific oyster family and belongs to the technical field of genomic molecular markers. The method comprises the following steps: (1) searching type I and type II SNP mutation sites in an SNP database acquired by an Illumina GoldenGate method; (2) establishing genotype database files of filial generations and parents, and acquiring genetic parameters of each SNP site by utilizing Cervus 3.0 software; (3) performing family simulation identification and analysis according to the allele frequency of each site by utilizing the Cervus 3.0 software, performing actual family identification and analysis, calculating a LOD value of a candidate parental pair, and selecting parents with the highest LOD value being positive value as real parents. According to the invention, an SNP marking method applied to identification of pacific oyster family is developed, and the reaction system and polymerase chain reaction (PCR) amplification conditions are optimized. The experiments prove that the primers have the characteristics of high repeatability, high stability, high solubility curve resolution and the like, and a foundation is laid for breeding of the pacific oyster family.

Description

technical field [0001] The invention belongs to the technical field of genome molecular markers, and relates to a method suitable for identification of Pacific oyster family. Background technique [0002] Oysters belong to the phylum Mollusca, Bivalvia, and Pearl Oyster. Its meat is delicious and nutritious. It has been eaten by humans since ancient times, and has high economic value and medicinal value. Oysters are a worldwide distribution group, and more than 100 species have been found so far. Almost all coastal countries in the world produce oysters. They are currently the economic shellfish with the largest output in my country and the world. [0003] As the world's largest cultured shellfish, the genetic improvement of oysters has always been widely valued by experts and scholars at home and abroad, and a lot of research work has been carried out. Although my country is a large oyster farming country, it is not yet a strong oyster farming country. There are problems s...

Claims

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Application Information

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IPC IPC(8): G06F19/24G06F19/28C12Q1/68
Inventor 孙秀俊杨爱国吴彪
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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