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Patinopecten yessoensis living body sex identification method based on DNA molecular markers

A DNA molecule and technology of Ezo scallop, applied in the field of live sex identification of Ezo scallop, can solve the problem that the method of sex identification of live Ezo scallop has not yet been established, and achieve the effect of high accuracy

Active Publication Date: 2019-12-24
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] So far, the sex identification method of live Ezo scallop has not been established

Method used

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  • Patinopecten yessoensis living body sex identification method based on DNA molecular markers
  • Patinopecten yessoensis living body sex identification method based on DNA molecular markers
  • Patinopecten yessoensis living body sex identification method based on DNA molecular markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Obtainment of sex-specific molecular markers of Ezo scallop

[0017] Genomic DNA of the adductor muscle of a female scallop was extracted by the classic phenol-chloroform method; sequenced by Pabio-Sequel sequencer at a depth of 100×; sequences were assembled using Falcon to obtain the genome of the female scallop; compared by mauve software In the genomes of female and male scallops, a sequence was found only in female individuals, but not in male individuals. This sequence is shown in SEQ ID NO:1.

Embodiment 2

[0018] Embodiment 2: living body detection method

[0019] Samples to be inspected: Take 25 Ezo scallops and collect 2 gill filaments for DNA extraction.

[0020] DNA extraction: Genomic DNA was extracted using the classic phenol-chloroform method, and the quality of the DNA was detected by agarose gel electrophoresis and a nucleic acid quantifier. figure 1 It is a schematic diagram of DNA electrophoresis results extracted from gill filaments obtained from living organisms. Numbers 1-16 in the figure are extracted DNA samples, and M indicates 1kb DNA ladder.

[0021] Primer synthesis: Design female-specific primers for amplification according to the obtained female-specific fragment SEQ ID NO:1. The specific primer sequences are shown in the table below.

[0022]

[0023] PCR amplification: The above primers were used for PCR amplification of the collected samples respectively. The reaction system is: DNA template 100ng, 5×HF buffer 4μL; dNTP (10mM) 0.6μL; upstream prime...

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Abstract

The invention provides a patinopecten yessoensis living body sex identification method based on DNA molecular markers. The patinopecten yessoensis living body sex identification method based on the DNA molecular markers comprises the steps that a small amount of gill filaments is obtained from the patinopecten yessoensis through a living body, DNA is extracted, PCR amplifies the sex-specific DNA molecular markers, whether a target band exists in a PCR product or not is detected by electrophoresis, and patinopecten yessoensis living body sex identification is achieved. The sex-specific DNA molecular markers are unique to female, and a nucleotide fragment sequence of the sex-specific DNA molecular markers is SEQ ID NO:1. PCR primers are used for amplifying the molecular markers, and the sexof patinopecten yessoensis is identified based on the agarose gel electrophoresis result; and a sample amplifying a 1859bp specific band is female, and samples without the band are male. The patinopecten yessoensis living body sex identification can be realized, the method is simple, the result is accurate, and the method is helpful to speed up the breeding process of the patinopecten yessoensis.

Description

technical field [0001] The invention belongs to the technical field of sex identification in aquaculture, and in particular relates to a method for sex identification of live scallops based on DNA molecular markers. Background technique [0002] Scallops are one of the most important maricultured shellfish in northern my country. At present, large-scale farming has been formed in the Bohai Sea and the northern Yellow Sea, creating an output value of billions of yuan in the past 10 years. Ezo scallops are dioecious, and generally reach sexual maturity in 1-2 years. When the gonads enter the growth and maturity stages, the sex of the scallops can be identified with the naked eye. However, when the gonads are in the undifferentiated, telogen, or proliferative stages, it is difficult to identify the sex with the naked eye, and it is necessary to dissect the gonads, and perform gender identification through histological sections or quantitative analysis of gene expression. Howev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6879C12N15/11
CPCC12Q1/6888C12Q1/6879
Inventor 张玲玲郭振义李亚娟刘亮洁王师陆维包振民
Owner OCEAN UNIV OF CHINA
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