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Method for selecting specific SRAP marking tapes for coilia ectenes genders and gender determination method

A technology of specific labeling and screening methods, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. Gender identification, etc.

Inactive Publication Date: 2013-01-30
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the dissection method has fatal damage to the fish body, and the external morphology observation can only be used to identify sexually mature broodstock during the breeding season
There is no effective method for gender identification of mature swordfish that have not reached sexual maturity or are not in the breeding season; the existing sex identification technology has seriously restricted the development of artificial propagation and pond culture technology of swordfish

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  • Method for selecting specific SRAP marking tapes for coilia ectenes genders and gender determination method
  • Method for selecting specific SRAP marking tapes for coilia ectenes genders and gender determination method

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Embodiment Construction

[0016] The present invention will be further described below in conjunction with the examples, but it does not constitute a limitation to the present invention.

[0017] 1. Screening of sex-specific SRAP marker bands

[0018] Including the following steps:

[0019] (1) Collect the samples of the anchovy, dissect the fish body for gender identification, and extract the total DNA of the female and male samples respectively;

[0020] (2) Design 14 SRAP primers, whose sequences are shown in Table 1;

[0021] Table 1 Primer information used in the SRAP-PCR reaction of the anchovy

[0022]

[0023] (3) Through preliminary experiments, explore the optimal SRAP-PCR reaction system: 25 μL of PCR system contains Mg 2+ 1.5 mmol / L, Taq enzyme 0.5U, template DNA 70ng, dNTPs 0.20mmol / L, primer 0.2μmol / L; amplification conditions are: 94°C for 3 min; 94°C for 1 min, 35°C for 1 min, 72°C for 1 min, 5 cycles; 94°C for 1 min, 50°C for 1 min, 72°C for 1 min, 35 cycles; 72°C for 8 min, st...

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Abstract

The invention discloses a method for selecting specific SRAP (Sequence-Related Amplified Polymorphism) marking tapes for coilia ectenes genders and a gender determination method. The specific SRAP marking tapes for coilia ectenes genders are selected through SRAP-PCR reaction of 49 primer sets; and then the DNA sample of the coilia ectene which is not clear in gender is amplified through a specific primer set; and the gender of the coilia ectenes is differentiated according to whether the corresponding specific marking tapes can be amplified or not. The method provided by the invention overcomes the limits and shortcomings of the existing coilia ectenes gender differentiation technology, and therefore, an accurate and convenient gender differentiation method is provided.

Description

technical field [0001] The invention relates to a screening method and a sex identification method for sex-specific SRAP marker bands of anchovy, belonging to the field of fishery biotechnology. Background technique [0002] Swordfish ( Coilia ectenes Jordan et Seale), also known as long-jawed anchovy, commonly known as saury, hair saury, belongs to Clupeiformes, Clupeidae, genus Coilia. Its meat is tender and delicious, and it is an important migratory economic fish in the middle and lower reaches of the Yangtze River. Due to many years of continuous high-intensity fishing and water environment pollution, the natural resources of sword anchovy have declined sharply, and the trend of younger migratory populations is obvious. In order to protect and utilize the resources of the cuttlefish, many scientific research institutes have begun to try pond culture and artificial propagation of cuttlefish in recent years. In order to realize the artificial breeding of sword anchovy,...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 张敏莹施炜纲徐东坡刘凯段金荣方第安
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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