Atlantic salmon DNA bar code standard detection gene, and primer and application thereof

A standard detection and barcode technology, applied in the field of molecular biology, can solve the problem that consumers cannot judge, judge its species, and have no form, and achieve the effect of shortening identification time, accurate identification and good stability.

Inactive Publication Date: 2021-04-16
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the differences in the meat quality of different species of salmon and trout, their breeding costs, sales prices and nutritional value are also different, and the consumption preferences of different countries are different, resulting in this phenomenon of confusing sales.
For the salmon sold in the market, most of them are muscles, bones and other tissues. There is no complete shape to judge its species, and consumers cannot judge it.

Method used

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  • Atlantic salmon DNA bar code standard detection gene, and primer and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The Atlantic salmon DNA barcode standard detection gene of this embodiment is shown in SEQ ID NO: 1 in the sequence listing.

[0022] Primers used to detect Atlantic salmon DNA barcode standard detection genes, including forward primer 5'-CTCTATTTTAGTATTTGGTGCCTGA-3' and reverse primer 5'-TAGACTTCTGGGTGGCCAAAGAACC-3'.

[0023] Application of the above Atlantic salmon DNA barcode standard detection gene in identification of Atlantic salmon. The specific identification methods are:

[0024] 1. Separation and extraction of DNA from the tissue samples to be tested;

[0025] 2. Use the DNA obtained in step 1 as a template, and use forward primers and reverse primers to perform PCR amplification; the conditions of the PCR amplification reaction are: 95°C pre-denaturation for 3min; 95°C denaturation for 15sec, 50°C annealing 10sec, 40sec extension at 72°C, a total of 35 cycles; the final extension was 7min at 72°C.

[0026] 3. Separate the PCR amplification product obtained...

Embodiment 2

[0028] 1) Detection sample DNA extraction:

[0029] Ten test samples were selected, among which S1, S2, and S3 were Atlantic salmon (Salmo salar) from Chile; S4, S5, and S6 were Atlantic salmon from Norway; S7 and S8 were rainbow trout (Oncorhynchus mykiss) from Norway; S9 was from the United States. Coho salmon (Oncorhynchus keta); S10 is Chilean coho salmon (Oncorhynchus kisutch). The above four fishes all belong to the family Salmonidae of the order Salmoniformes.

[0030] In this experiment, DNA was extracted according to the instructions of TIANampMarineAnimalsDNAKit Marine Biological Tissue Genomic DNA Extraction Kit (Spin Column Type) (Tiangen Biochemical Technology Co., Ltd.). Cut no more than 30mg of tissue material, put it into a centrifuge tube filled with 200μl GA buffer, vortex for 15sec; add 20μl ProteinaseK (20mg / ml) solution, vortex and mix well, and centrifuge briefly to remove water droplets on the inner wall of the tube cap ; Place at 56°C until the tissue...

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Abstract

The invention relates to an Atlantic salmon DNA bar code standard detection gene, and primers and application thereof, and belongs to the field of molecular biology. The invention also provides a pair of primers for detecting the Atlantic salmon DNA bar code standard detection gene, and application of an Atlantic salmon DNA bar code standard detection sequence in identifying Atlantic salmons. The primers are used for amplifying Atlantic salmon DNA, an obtained PCR amplification product is subjected to nucleotide sequence determination, a species identification result is judged according to a sequence comparison analysis result, and if the genetic distance between the PCR amplification product and a nucleotide sequence shown in SEQ ID NO: 1 is 0.02 or below, it can be judged that a sample to be detected is Atlantic salmon. According to the method, the Atlantic salmons can be distinguished from other salmons and trout types.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to an Atlantic salmon DNA barcode standard detection gene, a primer and an application thereof. Background technique [0002] Atlantic Salmon (Salmo salar) belongs to Salmoniformes (Salmoniformes) Salmonidae (Salmonidae) Salmon genus (Salmo), mainly distributed in the North Atlantic Ocean and in the rivers of northern Europe and the east coast of North America. Typical anadromous fish. They live in sea water most of the time, and during the breeding season, they swim to the river where they were born to lay eggs. The young fish will also grow up in the freshwater environment, and then return to the ocean to fatten up. Atlantic salmon is rich in unsaturated fatty acids and astaxanthin, and has extremely high nutritional and health value. It has become an internationally recognized high-end aquatic product and an excellent aquatic product with high development and breedin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
Inventor 柳淑芳赵新宁李昂牟铭庄志猛
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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