Identification on cods through loop-mediated isothermal amplification and primer used in identification

A loop-mediated isothermal, cod technology, applied in the field of temperature amplification detection, can solve problems such as identification of deep-processed cod products that are not used

Active Publication Date: 2019-07-16
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is currently no species identificat

Method used

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  • Identification on cods through loop-mediated isothermal amplification and primer used in identification
  • Identification on cods through loop-mediated isothermal amplification and primer used in identification
  • Identification on cods through loop-mediated isothermal amplification and primer used in identification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Embodiment 1, adopt two kinds of LAMP detection methods to carry out species identification to the cod (chilled or frozen) purchased on the market, comprise carrying out the following steps:

[0112] ① Extraction of genomic DNA in the sample to be tested; use animal tissue genomic DNA extraction kit (ZP307) (ZP307) to extract sample DNA and store at -20°C.

[0113] ② Species identification was carried out with the established detection system of 3 cod species (Atlantic cod, Pacific cod and haddock) and cod family LAMP detection system. details as follows:

[0114] 1. Real-time fluorescent LAMP amplification system setup: the reaction system is 10 μL, the system contains 5 μL of reaction buffer, 0.2 μL of upstream and downstream outer primers (10 μM, see Table 2), and 0.2 μL of upstream and downstream inner primers (50 μM, see Table 2). 0.32 μL, Bst polymerase 0.4 μL, 10 μM 9 fluorescent dye 0.2 μL, DNA template 0.5 μL, add ddH 2 0 to 10 μL;

[0115] Set up the syst...

Embodiment 2

[0137] Embodiment 2, adopt two kinds of LAMP detection methods as described in embodiment 1 to carry out species identification to the cod deep-processing commodity that buys randomly on the market:

[0138] Randomly purchased from the market marked as cod sausages, baked cod fillets and products containing cod in the product ingredients, using the animal tissue genomic DNA extraction kit (ZP307) to extract sample DNA, and stored at -20 °C.

[0139] That is, step ① in Embodiment 1 has been changed, and the rest of the content is equivalent to Embodiment 1.

[0140] Table 4. Species identification results of commercially available deep-processed cod products by LAMP detection method

[0141]

[0142]

[0143]

[0144]

[0145] As shown in Table 4, the 12S rDNA system in the 6 deep-processed samples was positive, which indeed belonged to cod. Atlantic cod and Pacific cod were detected at the same time in samples No. 2 and No. 6. Since the price of Pacific cod in the...

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Abstract

The invention discloses a primer for identification on cods through loop-mediated isothermal amplification. The primer comprises a primer in accordance with gadus morhua, a primer in accordance with gadus macrocephalus, a primer in accordance with melanogrammus aeglefinus and a primer in accordance with gadidae cods. The invention further provides a method for identifying the cod through loop-mediated isothermal amplification by the primer. The method comprises the following steps of (1), extracting genome DNA from samples (including fish samples) to be detected; (2) performing real-time fluorescence LAMP method and LAMP dyeing method through the primer; and (3) performing judgment according to the detection result obtained through the real-time fluorescence LAMP method and the LAMP dyeingmethod. The loop-mediated isothermal amplification is applied to variety identifying and marking research of common cods (the gadus morhua, the gadus macrocephalus and the melanogrammus aeglefinus) and relevant products common in domestic market.

Description

technical field [0001] The invention relates to a loop-mediated isothermal amplification detection technology for domestic cod fish products. Background technique [0002] Cod is mostly a cold-water fish that lives in the bottom of the ocean and the middle and lower layers of the deep sea, belonging to the phylum Vertebrata, the class Osteichthyes, and the order Gadiformes. Cod meat is luscious and nutritious. It is one of the main edible fish and occupies an important position in the world fishery. At present, the consumption of cod in my country is increasing year by year, and commercially available cod is mainly divided into three categories: (1) cod belonging to the cod family, including Atlantic cod (Gadus morhua) and Pacific cod (Gadus macrocephalus), Pollock (Theragra chalcogramma) and other fish of the cod family, such as haddock (Melanogrammus aeglefinus), pollock (Theragrachalcogramma), etc.; Anoplopoma fimbria) and Antarctic toothfish (Dissostichus eleginoides), w...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11C12Q1/6844
CPCC12Q1/6888C12Q1/6844C12Q2531/119C12Q2561/113C12Q2563/107Y02A40/81
Inventor 冯俊丽汪艺朱勤超沈清
Owner ZHEJIANG GONGSHANG UNIVERSITY
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