A SNP molecule mark for identifying individual gender of Takifugu rubripes and an identification method

A technology of redfin puffer and molecular markers, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem of low accuracy and achieve the effect of easy operation and improved accuracy

Inactive Publication Date: 2018-11-09
DALIAN OCEAN UNIV
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Problems solved by technology

Takashi Kamiya, et al., Plos Genetics, 2012, 8(7):e1002798, have confirmed that the 7271st locus of the Amhr2 gene of the red fin puffer is the sex determinati...

Method used

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  • A SNP molecule mark for identifying individual gender of Takifugu rubripes and an identification method

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Embodiment Construction

[0011] The identification method of the embodiment of the present invention is carried out according to the following steps:

[0012] a. extracting the genomic DNA of the redfin puffer sex individual to be identified;

[0013] a.1 Acquisition of puffer populations

[0014] The individuals used for gender identification were red-finned puffers from different families of puffers in different farms in Dalian, Hebei and other places. 252 individuals were randomly selected, and a few caudal fin rays were cut and placed in 80% ethanol at -20°C. Store in the refrigerator for extraction of genomic DNA.

[0015] a.2 Extraction of redfin puffer genome DNA

[0016] The genomic DNA in the fin rays of puffer puffer was extracted by the conventional phenol imitation method, and the specific steps were as follows:

[0017] (1) Take 100~120 mg of the caudal fin tissue of puffer puffer in a 1.5 ml centrifuge tube, and use ophthalmic scissors to cut it as much as possible (in powder form); ...

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Abstract

The invention discloses a SNP molecule mark for identifying individual gender of Takifugu rubripes. The SNP molecule mark for identifying individual gender of Takifugu rubripes is characterized in that a DNA sequence is shown in SEQ ID NO: 1, and a base N of the 121st position and the 262th position in the sequence is C or G. The identification method perform according to the following steps: a Takifugu rubripes genome DNA is used as a template, PCR amplification is performed by upstream primer and downstream primer, a PCR product is obtained; the DNA sequences of the upstream primer and the downstream primer is respectively shown in SEQ ID NO: 2 and SEQ ID NO: 3; sequencing is performed on the obtained PCR product, the individual that the 121st position and the 262th position of the measured DNA sequence are simultaneously double peaks is a Takifugu rubripes male fish, and the individual that the 121 position and the 262th position of the measured DNA sequence are simultaneously a single peak is a Takifugu rubripes female fish.

Description

technical field [0001] The invention relates to a SNP molecular marker and identification method for identifying the individual sex of red-finned puffer, especially a SNP for identifying the sex of red-finned puffer, which can simultaneously obtain male and female determination results, is easy to operate, and can improve the accuracy of identification Molecular markers and identification methods. Background technique [0002] Redfin Pufferfish ( Takifugu rubripes ) is an important sea-farming economic fish in the northern coast of my country. Its meat is delicious and nutritious, and it is favored by consumers in East Asian countries. The aquaculture output of redfin puffer per year in my country can reach more than 5,000 tons, and the artificial propagation, seedling raising, and growing technologies are basically mature, but there is still a big gap with the needs of the industry in terms of germplasm resources and new species creation. To find out the population resourc...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6879
CPCC12Q1/6879C12Q1/6888C12Q2600/156
Inventor 仇雪梅王智诚李鑫胡子文王秀利
Owner DALIAN OCEAN UNIV
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