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Molecular marker for distinguishing Heilongjiang megalobrama terminalis from southern megalobrama fishes, kit and distinguishing method

A molecular marker and kit technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as lack of intuitive and obvious differences, and achieve high accuracy and simple operation process. , the effect of short duration

Active Publication Date: 2018-07-13
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

Judging from the existing morphological studies, there is still a lack of intuitive and obvious differences to distinguish Heilongjiang triangular bream from southern bream fishes

Method used

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  • Molecular marker for distinguishing Heilongjiang megalobrama terminalis from southern megalobrama fishes, kit and distinguishing method
  • Molecular marker for distinguishing Heilongjiang megalobrama terminalis from southern megalobrama fishes, kit and distinguishing method
  • Molecular marker for distinguishing Heilongjiang megalobrama terminalis from southern megalobrama fishes, kit and distinguishing method

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specific Embodiment approach 1

[0023] Specific embodiment one: the present embodiment distinguishes the primer pair of molecular markers of Heilongjiang triangular bream and southern bream fishes as HLJFLf1 and HLJFLr1, and the specific sequences are:

[0024] HLJFLf1: 5′-GCATCTGGCTTCAATCTCA-3′

[0025] HLJFLrl: 5'-GATTTGCTGAGCGTAGGG-3'.

specific Embodiment approach 2

[0026] Specific embodiment 2: The application of molecular markers in this embodiment to distinguish Heilongjiang triangular bream from southern bream.

specific Embodiment approach 3

[0027] Specific embodiment three: the difference between this embodiment and specific embodiment one is: the specific method for molecular markers to distinguish Heilongjiang triangular bream and southern bream is as follows:

[0028] 1. Extract the genomic DNA of the fin rays of the bream fish to be tested for subsequent use;

[0029] 2. Using the extracted genomic DNA as a template and using HLJFLf1 and HLJFLr1 as primers for PCR amplification to obtain PCR products;

[0030] 3. Use the restriction endonuclease TaqI to digest the PCR product;

[0031] 4. Use 1.5% agarose gel to carry out electrophoresis detection on the digested products. The corresponding individual with 200bp and 180bp double bands in the electrophoresis results is Heilongjiang triangular bream, and the individual without 200bp and 180bp double bands is not Heilongjiang triangular bream. That is to say, the identification is completed;

[0032] Wherein the nucleotide sequence of HLJFLf1 is: 5'-GCATCTGGCT...

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Abstract

The invention discloses a molecular marker for distinguishing Heilongjiang megalobrama terminalis from southern megalobrama fishes, a kit and a distinguishing method, and relates to the molecular maker for distinguishing the megalobrama fishes and application of the molecular marker. A primer pair of the molecular marker can be HLJFLf1 and HLJFLr1. The kit comprises molecular marker primer pairs HLJFLf1 and HLJFLr1. The distinguishing method comprises the following steps: extracting fin rays DNA; 2, performing PCR amplification by using the HLJFLf1 and the HLJFLr1 as primers; 3, performing enzyme cutting on a PCR product by utilizing restriction endonuclease; 4, performing electrophoresis, wherein an electrophoresis result shows that corresponding individuals of double straps 200bp and 180bp are Heilongjiang megalobrama terminalis. According to the method disclosed by the invention, representative geographical population of the Heilongjiang megalobrama terminalis from the southern megalobrama fishes can be quickly, effectively and stably distinguished under a situation of no cross breeding.

Description

technical field [0001] The present invention relates to a molecular marker for distinguishing fishes of the genus Bream and its application. Background technique [0002] The existing bream fishes in my country include 4 species, which are pachygnathic bream in the upper reaches of the Yangtze River, grouphead bream (Wuchang fish) in large lakes in the middle and lower reaches of the Yangtze River, Guangdong bream in the Pearl River and various water systems of Hainan, and bream widely distributed in many water systems. Triangle bream. At present, there are 3 representative geographical populations of Trigonium bream, namely Qiantang River Triangle Bream, Yangtze River Triangle Bream and Heilongjiang Triangle Bream. Heilongjiang triangular bream is the "Faro" in the famous fish "San Hua Wu Luo" unique to the Heilongjiang River Basin, and it is also the only naturally distributed bream fish in the north of the Yellow River. As a local medium and large economically famous fis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2521/301C12Q2565/125
Inventor 胡雪松石连玉李池陶栾培贤曹春花贾智英尚梅葛彦龙王世会张秋军
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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