Method for rapidly identifying charybdis feriata sex specific molecular marker and genetic sex

Abstract

The invention relates to a method for rapidly identifying a charybdis feriata sex specific molecular marker and genetic sex. For SNP sites used for charybdis feriata sex identification, the base sequence is SEQ ID NO:1 or SEQ ID NO:2, and the length is 368bp; the SNP sites are the 35 site, the 89 site, the 120 site, the 218 site and the 240 site of SEQ ID NO:1 or SEQ ID NO:2. The identification method comprises the steps that charybdis feriata male and female individuals with known sex is subjected to DNA extraction; genomic library building and high throughput sequencing are simplified; sequence analysis and sex specific SNP site screening are conducted; candidate SNP site enlarged sample validation is conducted; sex specific primers based on the sex specific SNP sitesare designed; by means of the male specific primers, under specific annealing temperature, the genome DNA of charybdis feriata is subjected to PCR amplification, and specific sex determination is conducted according to the agarose gel electrophoresis detection result. A reliable method can be provided for charybdis feriata genetic sex and especially for the charybdis feriata in early developmentstage, and the method has the advantages of being easy to operate, short in consumed time, low in cost, good in repetition, high in accuracy rate and the like.

Description

technical field [0001] The invention belongs to the marine crab sex identification technology in the technical field of aquatic organisms, and in particular relates to a method for quickly identifying sex-specific molecular markers and genetic sex of rusty spot moth. Background technique [0002] SNP (Single Nucleotide Polymorphism) refers to the variation of a single nucleotide on the genome, including transitions, transversions, deletions, and insertions, forming genetic markers with a large number and rich polymorphisms. As the third generation genetic molecular marker, SNP has been widely used in the research of genetic diversity analysis, gene mapping, molecular marker-assisted breeding and functional genomics of many species. [0003] Charybdis feriatus, commonly known as safflower crab, belongs to the class Crustacea, order Decapoda, family Portunidae, genus Charybdis, and is mainly distributed in the tropical, subtropical and temperate regions of the Pacific Ocean an...

AI Technical Summary

Problems solved by technology

However, due to the fact that the technical means of screening polymorphic genetic markers through simplified genome sequencing is not widely used, and there is no combination of molecular methods for identifying the sex of rusty spot. Now, in production practice or academic research, only morphological methods can be used to identify rusty spot. Gender, the identification accuracy is low, and it is difficult to identify the sex of the rusty spot in the early stage of growth and development. There are no reports on sex-linked markers and genetic sex identification methods with the rusty spot.

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