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Method for quickly identifying oplegnathus punctatus and scatophagus argus

A technology of zebra sea bream and goldfish, which is applied in the directions of biochemical equipment and methods, determination/inspection of microorganisms, etc., to achieve the effects of accurate identification, low requirements for equipment and equipment, and low price

Active Publication Date: 2019-12-24
INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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  • Method for quickly identifying oplegnathus punctatus and scatophagus argus

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Embodiment 1

[0022] (1) Extract the genomic DNA of the grouper and money fish samples:

[0023] ①Use simple and sterilized scissors to cut off the grouper bream and money fish about 5mm 2 The caudal fin tissue was slightly crushed with tweezers and placed in a clean 1.5ml plastic centrifuge tube.

[0024] ② Add 600μl lysate, lysate includes 1) Tris-Hcl 10molL -1 , EDTA-2Na 1mol L -1 , pH=8.0; 2) 10% SDS; 3) Proteinase K 20mgmL -1 ; 1), 2), 3) the volume ratio is 20:10:1;

[0025] ③ Add 600 μL of Tris saturated phenol:chloroform:isoamyl alcohol (25:24:1) after 3 hours in a water bath at 55°C, shake gently for 20 minutes, then centrifuge at 5000 r / min for 10 minutes, repeat once;

[0026] ④ Transfer the extracted supernatant to a clean 1.5mL centrifuge tube, pour 2 times of absolute ethanol (pre-cooled at -20°C), and centrifuge at 12000r / min for 10min;

[0027] ⑤Pour out absolute ethanol, pour 1mL of 75% ethanol to dissolve DNA, centrifuge at 12 000r / min for 5min, repeat once;

[0028]...

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Abstract

The invention relates to a method for quickly identifying oplegnathus punctatus and scatophagus argus. The method comprises the steps of (1) extracting genome DNA from oplegnathus punctatus samples and scatophagus argus samples; (2) performing PCR amplification through a specific primer; and (3) performing gel coagulation and electrophoresis on PCR amplification products, taking a photograph, andrecording a comparison electrophoresis result, wherein the oplegnathus punctatus has a clear specificity identifying strip at 160bp, and the scatophagus argus has a clear specificity identifying stripat 200bp. The method disclosed by the invention is accurate, simple and quick to identify, low in price, low in requirements for instruments and equipment, suitable for large-scale identification operations and has favorable application prospects.

Description

technical field [0001] The invention belongs to the field of fish identification, in particular to a method for rapidly identifying porpoise and money fish. Background technique [0002] Spotted seabream (Oplegnathus punctatus) belongs to the genus Oplegnathus punctatus of the family Perciformes, commonly known as spotted seabream, black mouth, flower golden drum, etc. Since the grouper is mainly distributed in the Pacific Ocean, there are few natural resources. There are only a small number of units and breeding companies in my country for tentative breeding of bream, and the scale of production has not yet formed. Spotted sea bream has high edible and medicinal value, delicate meat, rich collagen, unique taste, rich in various amino acids and unsaturated fatty acids, and has the reputation of "sashimi masterpiece" in Japanese cuisine. In southern Fujian, porphyra is a species that can be encountered but not sought after. Fishermen often catch porpoise off the coast of Pe...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/6888C12Q1/686C12Q2565/125Y02A40/81
Inventor 祝斐吴国均张曹进陈淑吟贾超峰孟乾张志伟高波孙瑞建沙士兵於明伟陈自强曾海峰林士杰
Owner INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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