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SNP sites for sex determination of Scylla paramamosain and identification method thereof

An identification method and technology for Scylla simulacrum, which are applied in the fields of biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, etc. and other issues, to achieve the effect of wide promotion and application potential, strong versatility and high application value

Active Publication Date: 2017-07-11
SHANTOU UNIV
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AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a SNP site and its identification method for the sex identification of Scylla pseudocaveus, so as to solve the problems that the sex of Scylla pseudocavena in the early stage of development cannot be identified, and the genetic sex of Scylla pseudocavena cannot be identified.

Method used

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Embodiment 1

[0025] 1. Extraction of genomic DNA of male and female individuals of Scylla pseudocavena.

[0026] First, 115 adult crabs, including 63 females and 52 males, were collected. Cut about 10 g of muscle tissue from gaiters, and homogenize it in an EP tube containing 300 µL of lysate; after that, add proteinase K at a final concentration of 25 µg / µL to digest the muscle tissue, and at the same time add a final concentration of 120 µg / µL of RNase A digested the RNA in the muscle tissue, and incubated it in a water bath at 50°C for about 2 h; finally, extracted twice with phenol and chloroform, precipitated the DNA with ice ethanol, and dried the DNA in a 37°C incubator. And dissolved in sterile double-distilled water, stored at low temperature for later use.

[0027] 2. Simplify the construction and high-throughput sequencing of genomic libraries.

[0028] In this study, simplified genome libraries of 10 female crabs and 10 male crabs were constructed. First, use EcoRI-HF endon...

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PUM

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Abstract

The invention relates to SNP sites for sex determination of Scylla paramamosain and an identification method thereof. A base sequence is SEQ ID No. 1 or SEQ ID No. 2 and has a length of 285 bp; and the SNP sites are site 62, site 70, site 74, site 145, site 147, site 163, site 186 and site 194 in the SEQ ID No. 1 or SEQ ID No. 2. The identification method comprises the following steps: extraction of the genome DNAs of male and female individuals; construction of a reduced genome library and high-flux sequencing; sequence analysis and screening of candidate SNP markers linked with sexes; and large-sample verification of the candidate SNP markers. According to the invention, the sex-linked molecular markers of Scylla paramamosain are identified for the first time and have great guidance significance to the sex determination and differentiation mechanism of Scylla paramamosain and to research on unisexual breeding. The method provided by the invention has the advantages of accuracy, sensitivity, reliability and the like, has good universality, does not need to learn about genome information in advance and is applicable to a great number of species without reference genomes, so the method has good promotion and application potential.

Description

technical field [0001] The invention belongs to the marine crab sex-linked molecular marker screening and genetic sex identification technology in the technical field of aquatic organisms, and in particular relates to a SNP site and an identification method for the sex identification of the mud crab. Background technique [0002] Decapod is a higher group of crustaceans, including many economically important shrimps and crabs, such as: Litopenaeus vannamei, Penaeus sinensis, Penaeus monodon, Eriocheir sinensis, Portunus trituberculatus, and mud crabs. The genus Scylla includes 4 species: Scylla serrata, Scylla elven, Scylla purple claw, and Scylla pseudocave. They are widely distributed in the Western Pacific, Southeast Asia, Australia, the Indian Ocean, and the coasts of Africa. One of the tallest saltwater species. Among them, the blue crab is mainly distributed in the coasts of the East my country Sea and the South China Sea, and is a traditional marine fishery resource ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 马洪雨苗贵东游翠红杨小龙程银伟章跃陵石西
Owner SHANTOU UNIV
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