Molecular markers and methods for identifying Zhuxing three-piece fish and three-piece fish

A technique for molecular labeling and identification of beads, which is applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., and can solve the problems of expensive capillary electrophoresis analysis process, cumbersome effective analysis and identification of alleles, and difficult implementation Problems such as complex process, to achieve the effect of protecting germplasm resources, fast identification process and strong operability

Active Publication Date: 2021-12-14
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The implementation process is relatively complex and time-consuming, the capillary electrophoresis analysis process is relatively expensive, and the effective analysis and identification of the amplified alleles is relatively cumbersome

Method used

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  • Molecular markers and methods for identifying Zhuxing three-piece fish and three-piece fish
  • Molecular markers and methods for identifying Zhuxing three-piece fish and three-piece fish

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specific Embodiment approach 1

[0020] Specific embodiment 1: In this embodiment, the first molecular marker primer pair for identifying Zhuxing three-piece fish and three-piece fish is A8, and the nucleotide sequence of the A8 forward primer is 5'-GGCTACAGCAAGGCAAGAAC-3'; the A8 reverse primer The nucleotide sequence is 5'-ATGCATTTCCCTTCGTATGC-3'.

specific Embodiment approach 2

[0021] Specific embodiment 2: The second molecular marker primer pair for identifying Zhuxing three-piece fish and three-piece fish in this embodiment is A11, and the nucleotide sequence of the A11 forward primer is 5'-GGACTGGGGAGGTGTGAGTA-3'; the A11 reverse primer The nucleotide sequence is 5'-TGCAGAAAGAGCAGCAGAAA-3'.

specific Embodiment approach 3

[0022] Specific implementation mode three: the method for identification of Zhuxing three-piece fish and three-piece fish in this embodiment:

[0023] 1. Extraction of sample DNA;

[0024] 2. PCR detection:

[0025] The PCR reaction system is 25 μl, consisting of 18 μl self-made PCR buffer, 1.0 μl forward detection primer with a concentration of 10 μmol / L, 1.0 μl reverse detection primer with a concentration of 10 μmol / L, 2 μl sample DNA, and 0.2 μl Taq with an enzyme activity of 5 U DNA polymerase and the rest of deionized sterile water; 18 μl self-made PCR buffer containing 50mmol / L of KCl, 10mmol / L of Tris-HCl, 0.1% volume of TritonX-100, 1.5mmol / L of MgCl 2 , 0.1% by volume of NP-40, 0.01% by volume of gelatin and 200 μmol / L of four kinds of d NTP mixture; wherein the detection primers are molecular marker primer pair A8 or molecular marker primer pair A11;

[0026] The amplification conditions of PCR are: pre-denaturation at 95°C for 3min, denaturation at 95°C for 15s, ...

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Abstract

Molecular markers and methods for identifying Zhuxing Yaluo and Sankuaiyu relate to the field of identification of Sankuaiyu genus populations. The first identification molecular marker primer pair of the present invention is A8. The second identification molecular marker primer pair of the present invention is A11. The method for identification of Zhuxing Yaluo and Sankuai: 1. Extraction of sample DNA; 2. PCR detection; 3. Agarose gel electrophoresis of PCR amplification products; 4. Judgment of PCR results. The method of the present invention simplifies the molecular identification steps of Zhuxing three-piece fish and three-piece fish germplasm, and can visually distinguish Zhuxing three-piece fish and three-piece fish according to the results of agarose gel electrophoresis, and has simple steps and stronger operability , The advantages of lower equipment requirements, faster identification process and more accurate identification results. The method of the invention can avoid germ plasm mixing caused by artificial propagation and protect germ plasm resources.

Description

technical field [0001] The invention relates to the field of group identification of the genus Sankuai. Background technique [0002] The genus Tribolodon is mainly distributed in the Sea of ​​Japan and some inland freshwater rivers in the northwest of the Pacific Ocean, including Russia's Primorsky Province, Khabarovsk and Sakhalin, Japan and the Korean Peninsula. In China, it is only distributed in Suifenhe and Tumen River Basin. Suifenhe is one of the main rivers in the Sea of ​​Japan. After the gonads of the three-piece fish are mature, they go up the river in batches and enter the estuary. They spawn in the water at the sharp beach sentry, hence the name "beachhead fish". Return to the offshore area immediately. According to the time of migration in batches and the difference in marriage color, local fishermen divide it into "Golden Tantou", "Silver Tantou" and "Black Tantou". The fertilized eggs are hatched in freshwater rivers, and the fry flow downstream to offsho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 苏宝锋赵雪飞常玉梅梁利群孙博王维坤
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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