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A Molecular Marker and Its Primer and Application for Distinguishing Common Carp, Crucian Carp and Crossbreed Fish between Carp and Crucian Carp

A technology for molecular marker and identification of carp, applied in the field of molecular genetics of aquatic animals, can solve the problems of difficult identification and differentiation, unclear background source, damaged processing form, etc., and achieves the effects of strong specificity, high amplification efficiency and low cost.

Active Publication Date: 2021-12-07
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the difficulty of identification and distinction caused by the existing morphological identification methods, such as the difficulty of distinguishing appearance, unclear background source, and damage to processing form, and the inability of mitochondrial DNA to solve the identification problems of carp, crucian carp and hybrid fish between carp and crucian carp

Method used

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  • A Molecular Marker and Its Primer and Application for Distinguishing Common Carp, Crucian Carp and Crossbreed Fish between Carp and Crucian Carp
  • A Molecular Marker and Its Primer and Application for Distinguishing Common Carp, Crucian Carp and Crossbreed Fish between Carp and Crucian Carp

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Embodiment 1

[0026] According to the published genome data information of carp and crucian carp in the NCBI database (National Center for Biotechnology Information). Carp and crucian carp genomes were analyzed and compared for homologous sequences. A large number of differential gene sequences were obtained by screening. Design primers from the homologous regions of these sequences, use the DNA of carp, crucian carp, and carp-crucian hybrid fish as templates for PCR amplification and product sequencing analysis, and screen and verify the differential sequences of carp, crucian carp, and carp-crucian hybrid fish , that is, the molecular marker, its nucleotide sequence is as SEQ ID NO.3 and SEQ ID NO.4, and the primer pair SEQ ID NO.1 and SEQ ID NO.2 corresponding to the molecular marker are also screened.

[0027] The above-mentioned molecular markers can be used to identify carp, crucian carp and hybrid fish between carp and crucian carp. The sample with the nucleotide sequence shown in S...

Embodiment 2

[0033] The present embodiment provides a kind of method for distinguishing between carp, crucian carp and carp and crucian carp hybrid fish, according to the following steps:

[0034] 1) extract the genomic DNA of the fish sample to be tested;

[0035] 2) using the genomic DNA of the fish sample to be tested as a template, using the primer pair shown in SEQ ID NO.1-2 to perform a PCR amplification reaction to obtain a PCR product;

[0036] 3) Electrophoresis detection of PCR products, if only a single fragment with a length of 481bp is obtained, the fish to be tested is carp; if only a single fragment with a length of 958bp is obtained, the fish to be tested is crucian carp; fragments, the fish to be tested is a hybrid between carp and crucian carp.

Embodiment 3

[0038] In this embodiment, 4 carp samples (respectively labeled samples 1-4), 4 crucian carp samples (respectively labeled samples 5-8) and 4 carp and crucian carp hybrid fish samples (respectively labeled samples 9-12) are used as samples to be tested Utilize respectively the primer pair and the identification method of the molecular marker of embodiment 1 to carry out identification and the accuracy of verification method on the gene level to carp, crucian carp and carp and crucian carp hybrid fish sample, concrete testing method and flow process are as follows:

[0039] 1. Genomic DNA extraction of the fish to be tested:

[0040] (1) Cut a small amount of fin rays or muscle tissue of the fish to be tested, and put them into a 2mL centrifuge tube;

[0041] (2) Add 600 μL DNA extraction lysate. Lysis solution components: 1M Tris pH8.0, 5M NaCl, 0.5M EDTA pH8.0, 10% SDS and 200μg / mL Protein K, mix well;

[0042] (3) digest at 55°C for 10 hours;

[0043] (4) Add an equal vol...

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Abstract

The invention discloses a molecular marker for identifying carp, crucian carp and hybrid fish between carp and carp, as well as its primer and application, belonging to the field of molecular genetics of aquatic animals. The nucleotide sequence of the molecular marker of the present invention is shown in SEQ ID NO.3 and SEQ ID NO.4. The nucleotide sequence of the molecularly marked primer pair is shown in SEQ ID NO.1‑2. The invention also provides a kit containing the primer pair of the molecular marker. And the application of the primer pair of the molecular marker in the identification of carp, crucian carp and hybrid fish between carp and crucian carp. The present invention develops a molecular marker based on carp genome knowledge, and provides a method for identifying carp, crucian carp, and carp-crucian hybrid fish at the genetic level, effectively avoiding indistinguishable appearance, unclear background sources, and processed forms Difficult to identify and distinguish problems caused by damage, etc. It has the advantages of simplicity, speed, efficiency and accuracy.

Description

technical field [0001] The invention relates to a molecular marker for identifying carp, crucian carp and hybrid fish between carp and carp, as well as its primer and application, belonging to the field of molecular genetics of aquatic animals. Background technique [0002] Carp (Cyprinus carpio) and crucian carp (Carassius auratus) are common and important freshwater economic fishes. At present, the annual output in my country is more than 3 million tons. Carp (Cyprinus carpio) and crucian carp (Carassius auratus) both belong to Cyprinidae, Cyprinidae, and have close relationship and similar morphological structure. Carp grows faster and matures later than crucian carp, and the size of commercial fish is usually 1-2 kg; crucian carp grows slower than carp, usually reaches sexual maturity in one year, and the size of commercial fish is usually about 0.5 kg. Although carp has more advantages in terms of growth speed and other aspects, crucian carp meat is more tender and del...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 程磊王乐李超鲁翠云
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI