Specific primer and method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis

A technology of red-finned puffer and puffer obscura, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of high cost, cumbersome operation, time-consuming, etc., and achieve low cost , Simple operation, high precision effect

Active Publication Date: 2020-11-24
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The operation is cumbersome

Method used

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  • Specific primer and method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis
  • Specific primer and method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis
  • Specific primer and method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Obtaining samples from three populations of puffer obscurus, puffer red fin and hybrid puffer

[0033] The materials collected included 24 samples of eight confirmed species of puffer, red-finned puffer and hybrid puffer, among which puffer seedlings came from Jiangsu Zhongyang Group Co., Ltd. The Lukou Experimental Base of the Institute of Fishery Sciences breeds 5-month-old puffers and collects materials; 5-month-old pufferfish and puffer hybrids are all obtained from Jiangsu Zhongyang Group Co., Ltd. (Haian Base).

[0034] (2) Extraction of genomic DNA from puffer obscurus, puffer red fin and hybrid puffer

[0035] 10-30 mg of caudal fin was taken from each fish, and genomic DNA was extracted from 24 puffer fish by using a DNA extraction kit (purchased from Beijing Baitaike Biological Company).

[0036] (3) Screening of microsatellite loci and synthesis of primers

[0037] Firstly, the microsatellite screening software MISA was used to screen the microsatellite...

Embodiment 2

[0052] (1) Obtaining different groups of Fugu obscurus, Fugu red fin and hybrid Fugu:

[0053] The materials collected include 60 samples of 20 puffers obscurus, 20 puffers obscurus and 20 hybrid fugus hybrids, among which 20 puffers obscurus 5 months old were from Jiangsu Zhongyang Group Co., Ltd. (Haian, 8 ), Zhenjiang Jiangzhiyuan Fishery Technology Co., Ltd. (Yangzhong, 6 fishes) and Jiangyin Shengang Sanxian Breeding Co., Ltd. (Jiangyin, 6 fishes); 20 3-year-old wild redfin pufferfish were taken from Shidao, Yantai, Shandong; 20 5 fishes Month-old cultured hybrid oriental puffer was obtained from Jiangsu Zhongyang Group Co., Ltd. (Haian base).

[0054] (2) Extraction of genomic DNA from puffer obscurus, puffer red fin and hybrid puffer

[0055] For specific genomic DNA extraction steps, refer to step (1) in Example 1.

[0056] (3) Analysis of experimental results of puffer obscurus, puffer red fin and hybrid puffer

[0057] For specific experimental steps, refer to Exa...

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Abstract

The invention discloses a specific primer and method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis. The specific primer comprises a forward primer and a reverseprimer, and nucleotide sequences of the forward primer and the reverse primer are respectively shown as SEQ ID NO.1-2. The identification method comprises the following steps of firstly, collecting apuffer fish sample to be identified, and extracting genome DNA; then taking the genomic DNAs of the three puffer fishes as templates, and performing PCR amplification on the templates by using the primer; and finally, performing electrophoresis on amplification products and performing identifying according to strip difference. The method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis by using the specific primer does not need complex steps such as microsatellite computer sequencing, can quickly and accurately distinguish the three kinds of fishes only according to the number and positions of strips, and has the advantages of good stability, strong practicability, simplicity in operation, high accuracy and the like.

Description

technical field [0001] The invention belongs to the technical field of aquaculture, and in particular relates to a specific primer and a method for identifying puffer puffer obscurus, puffer red fin and hybrid puffer. Background technique [0002] Takifugu fasciatus and Takifugu rubripes, commonly known as puffers, belong to Osteichthyes, Actinopterygii, Letrodontiforms, Tetraodontidae ), the genus Takifugu, is mainly distributed in the Yellow Sea, Bohai Sea and offshore waters, rivers, seas and lakes in China. The meat of the two is fresh, tender and plump, with high nutritional value. They are deeply loved by consumers and are aquatic products with high economic value. [0003] By recombining the genotypes of the parents, hybrid breeding enables the offspring to have the excellent traits of the parents, and may even have some shape characteristics that the parents do not have, and have certain heterosis. The hybrid puffer produced by the crossbreeding of puffer obscura f...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 尹绍武徐杰杰王涛李杰孙亦如朱永祥
Owner NANJING NORMAL UNIVERSITY
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