A method for identification of hybrid seabream families using microsatellite markers

A microsatellite marker, seabream family technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of no hybrid seabream family identification method, etc., and achieve the effect of strong stress resistance

Active Publication Date: 2021-07-02
INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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Problems solved by technology

So far, there is no method for identification of hybrid seabream families

Method used

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  • A method for identification of hybrid seabream families using microsatellite markers
  • A method for identification of hybrid seabream families using microsatellite markers
  • A method for identification of hybrid seabream families using microsatellite markers

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Embodiment 1

[0039] In April 2014, 10 red sea bream females (denoted as P R 1-P R 10) and 260 black sea bream Jiangsu populations to select 12 black sea bream males with mature gonads (denoted as P B 1-P B 12), according to the method of diallell hybridization, 120 families were established through artificial insemination. After the hybrid individuals grew to 90 days, the caudal fin rays were cut and stored in 100% alcohol, and 20 hybrid breams were Generational individuals (denoted as HF1-HF20) were identified for genetic relationship.

[0040]A method utilizing microsatellite markers to identify a hybrid seabream family, comprising the steps of:

[0041] (1) Cut the tail fin samples of the female red sea bream and the male black sea bream and their hybrid offspring, and extract the genomic DNA;

[0042] 1) Take 5-20 mg of fish fin tissue, cut it into pieces, put it in a 1.5 mL centrifuge tube, add 400 μL of tissue extract and 10 μL of proteinase K, shake and mix on a shaker for 1 min...

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Abstract

The invention relates to a method for using microsatellite markers to identify hybrid seabream families, belonging to the field of molecular markers. Firstly, the tail fin samples of the female red sea bream and male black sea bream and their hybrid offspring were cut, and the DNA was extracted, and then the DNA of the parents and the hybrid offspring were used as templates, and microsatellite marker primers were used for multiplex PCR amplification; The products were subjected to polyacrylamide gel electrophoresis, analyzed with BandScan5.0 software, combined with the length of PCR products to determine the size of alleles, and determined individual genotypes, and established genotype data files of hybrid seabream and parents according to the analysis results; using Cervus3.0 software Perform family identification. The present invention adopts microsatellite marker technology to distinguish male and female parents and offspring genotypes for hybridization on 10 microsatellite loci, can accurately and conveniently determine the family source of offspring of hybrid sea bream, and quickly and efficiently select and breed excellent hybrid sea bream family, It brings great convenience to genetic breeding work.

Description

technical field [0001] The invention relates to a method for using microsatellite markers to identify hybrid sea bream families, and belongs to the technical field of molecular markers. Background technique [0002] Black sea bream (Acantho pagrusschlegelii) is oblong in shape with flat sides, elegant in body, bright in color and delicious in meat quality. It is a rare and valuable marine fish. Black sea bream has strong stress resistance and a wide range of adaptability to temperature and salinity, but it has a long breeding cycle and poor disease resistance. In recent years, due to the lack of breeding and improvement of breeding fish, germplasm resources have begun to decline, diseases have become more and more serious, quality has declined, and economic benefits have declined significantly, which has affected the stability and sustainable development of black sea bream aquaculture. Red sea bream (Pagrosomus major) is a warm-temperate bottom carnivorous fish in coastal w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 张志伟张志勇贾超峰祝斐孟乾曹广勇陈自强曾海峰
Owner INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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