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Primer for nile tilapia germ plasm purity identification and PCR identification method

A Nile tilapia and identification method technology, applied in the field of biological detection and identification, can solve the problems of reducing economic benefits, difficulties, and large differences in pool specifications, and achieve the effects of avoiding unstable results, accurate results, and quick operation

Inactive Publication Date: 2016-03-09
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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AI Technical Summary

Problems solved by technology

Due to the large overlap of morphological characteristics between different species of tilapia, it is very difficult to distinguish different tilapia species based on their morphology alone.
In addition, it is easy to hybridize between tilapia varieties, and the hybrids are fertile, and the hybrids are similar in shape to their parents. Therefore, it is easy to cause tilapia germplasm to mix, leading to the degradation of good economic traits, which is currently restricting our country. Prominent Problems in the Development of Tilapia Aquaculture Industry
During the breeding process, the growth of male and female tilapias is significantly different, and the male fish grows faster than the female fish, which often leads to a large difference in the size of the pond, thereby reducing the quality of commercial fish and reducing economic benefits.

Method used

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  • Primer for nile tilapia germ plasm purity identification and PCR identification method
  • Primer for nile tilapia germ plasm purity identification and PCR identification method

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Step 1. Primer Design

[0021] According to the MyoD gene sequence of Nile tilapia, the primer design software Primer5.0 was used to design primers. The primer sequence is:

[0022] Forward: 5'-CATGGTGAGTTCAAGGAGCAC-3'

[0023] Reverse: 5'-GAGAAAGTTACACACACAGGCAGCT-3'.

[0024] Step 2. Genomic DNA Extraction

[0025] (1) Take 19 samples to be inspected, all of which were collected by the Yixing Fishery of the Freshwater Fishery Research Center of the Chinese Academy of Fishery Sciences;

[0026] (2) Blood was collected from the tail vein, using the AxyPrep Blood Genomic DNA Mini Kit produced by Axygen, and the genomic DNA was extracted according to the instructions.

[0027] Step 3, establishment of PCR identification method

[0028] (1) The reaction system of PCR is: 5 μL of 10× reaction MIX solution, 0.25 μL of forward and reverse primers, 1 μL of DNA template, make up to 10 μL with sterilized double distilled water;

[0029] (2) The reaction program of PCR is: p...

Embodiment 2

[0034] The germplasm purity of Nile tilapia was identified by the steps described in Example 1. Among them, 23 Nile tilapias to be inspected were collected from the Yixing fishery of the Freshwater Fisheries Research Center of the Chinese Academy of Fishery Sciences. Such as figure 2 As shown in the electrophoresis test, it was found that 23 fish to be tested had a 229bp band, all of which were pure Nile tilapia.

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Abstract

The invention discloses a primer for nile tilapia germ plasm purity identification and a PCR identification method. The primer is capable of amplifying an MyoD (Myogenic Differentiation Antigen) gene of nile tilapia, so as to generate a strip of 229bp specific amplification fragment. (1), the primer provided by the invention is used to amplify the genomic DNA of the nile tilapia, so as to generate the strip of 229bp specific amplification fragment, and a strip of 264bp band of blue tilapia, wherein two strips of hybrid tilapia are respectively 229bp and 264bp bands, which can be obviously distinguished, and thus the germ plasm purity identification of the nile tilapia can be simply and accurately carried out; (2), the specificity of the primer provided by the invention is high, and other fragments except the target fragment cannot be amplified; (3), the invention discloses the PCR identification method using the primer, and the method is fast and convenient in operation and accurate in result, so that the defect of unstable result brought about by adopting a conventional morphological method to identify the nile tilapia of different species is avoided.

Description

technical field [0001] The invention relates to a biological detection and identification technology, in particular to a primer for performing PCR identification on the germplasm purity of Nile tilapia and a PCR identification method using the primer. Background technique [0002] Nile tilapia belongs to Perciformes, Cichlidae, and Tilapia genus. It is native to Lake Tanganyika in Jordan. It was introduced and promoted in my country from Thailand in 1978. Now my country is the largest tilapia in the world. Non-fish producing and exporting countries. The main species of tilapia cultured in my country are Nile tilapia, Oria tilapia and Oni hybrid tilapia. Due to the large overlap of morphological characteristics among different species of tilapia, it is very difficult to distinguish different tilapia species only based on their morphology. In addition, it is easy to hybridize between tilapia varieties, and the hybrids are fertile, and the hybrids are similar in shape to their...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 李红霞俞菊华唐永凯李建林于凡何杰
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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