A method for screening long oyster broodstock with high content of unsaturated fatty acid c20:3ω6
A technology of unsaturated fatty acids and long oysters, applied in the field of genetic engineering and genetic breeding, can solve the problems of the small number of markers and the inability to understand the mechanism of genetic regulation
Active Publication Date: 2021-05-28
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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However, this method is based on prior information and develops polymorphic sites on known functional genes. The relative number of markers is small, and the main effect sites may not be obtained, and the genetic regulation mechanism of the phenotype cannot be determined. have a comprehensive understanding
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Embodiment 1
[0040] a) Collection of samples: Collect 100 individuals from wild populations in Jiaonan where seedlings were hatched at the same time, dissect them, take adductor muscle and remaining tissues, and store them at -80°C after quick-freezing with liquid nitrogen.
[0041] b) Extraction of DNA: extract the genomic DNA of 100 samples and measure the concentration by ultraviolet absorbance photometry, and dilute the genomic DNA to 10-20ng / uL with sterile water according to the measured concentration;
[0042] c) Using SnaPshot to detect the genotype of the SNP site: (1) Using specific primers to perform peripheral amplification. Take the above long oyster genomic DNA as a template, and use primers F and R to prepare a reaction system: 1uL of genomic DNA, 5uL of general PCR mix, 0.2uL of primers F and R, 3.6uL of sterilized double-distilled water; the reaction system can be scaled up year-on-year; The primer sequences are:
[0043] Upstream primer F: 5'-ACGTCACAAGTAGGTGTTTAGAGA-3' ...
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The invention relates to the development of a SNP marker for identifying individuals with a high unsaturated fatty acid eicosatrienoic acid (C20:3Ω6) content in oyster and an application method thereof. Through previous genome-wide association analysis, 38 clustered SNP signals located on chromosome 1 of oyster longum that were significantly correlated with C20:3Ω6 content were identified. The present invention develops a corresponding detection method for a SNP site at the scaffold460_104,049 base in this region, the site has two base forms of A / T, and the nucleotide sequence of 500bp upstream and downstream is as shown in SEQ ID No.1 shown. Through this detection method, the parent shellfish of AT genotype can be screened to guide oyster breeding. The invention provides a method for genotype identification of parent shellfish before breeding of seedlings to increase the content of unsaturated fatty acids in progeny.
Description
technical field [0001] The invention belongs to the field of genetic engineering and genetic breeding, and relates to a method for screening oyster broodstock with high unsaturated fatty acid C20:3Ω6 content and a pair of primers for related SNPs. Background technique [0002] Oysters are an important marine aquatic resource and one of the most important marine cultured shellfish in the world. Although my country's oyster production is very high, the export volume only accounts for 3.3% of the world's oyster trade volume, and the average price is only 1 / 3 of other countries' oyster products, which shows that most oysters produced in China can only be produced and sold by themselves, and it is difficult to enter the international market. market and domestic high-end market. The main reason why Chinese oysters are hard to find in the international market is that the product specifications do not meet the quality standards of international trade. In fact, this is also the main ...
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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/172
Inventor 张国范李莉孟杰宋凯李春燕史瑞辉刘圣刘步苏
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI