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Molecular identification primer, kit and method for five groupers

A grouper and kit technology, applied in the field of molecular biology, can solve the problems of inconvenient screening and management of fry, inapplicable molecular identification and differentiation, etc., and achieve the effects of scientific management, stable reaction and high detection accuracy

Active Publication Date: 2020-09-01
SOUTH CHINA AGRI UNIV
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AI Technical Summary

Problems solved by technology

Because the red-spotted grouper, oblique-banded grouper, Yunlong hybrid spot, tiger-dragon hybrid spot and red dragon hybrid spot have many similarities in morphological characteristics, especially in the fry stage, it is difficult to distinguish them with the naked eye , which brings a lot of inconvenience to the fry selection and management of the fry farm
CN108588241A discloses a molecular-specific marker primer and method for identifying brown-spotted grouper, but the primer is only suitable for identifying brown-spotted grouper, not for hybrid F1 of brown-spotted grouper and other groupers Molecular characterization of offspring, and other groupers

Method used

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  • Molecular identification primer, kit and method for five groupers
  • Molecular identification primer, kit and method for five groupers
  • Molecular identification primer, kit and method for five groupers

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Embodiment 1

[0038] Example 1 A molecular identification method for five species of grouper

[0039] A molecular identification method for five species of grouper, including the following steps:

[0040] (1) DNA extraction

[0041] Collect the fin rays of the grouper sample to be tested, and extract its genomic DNA by DNA extraction.

[0042] (2) PCR amplification

[0043] Perform PCR amplification on the genomic DNA of the grouper sample to be tested obtained in step (1) using primers with nucleotide sequences as shown in SEQ ID NOs: 1 to 2 according to the following PCR reaction system and PCR reaction procedure;

[0044] Upstream primer (SEQ ID NO:1): 5, -TCGCCTGTTTATCAAAAACATCGC-3,;

[0045] Downstream primer (SEQ ID NO: 2): 5'-TCCGGTCTGAACTCAGATCACGTA-3'.

[0046] The PCR reaction system is (short centrifugation after mixing):

[0047]

[0048] The PCR reaction procedure is: 95°C pre-denaturation for 5 minutes; 95°C denaturation for 30 seconds, 55°C annealing for 30 seconds, 72°C extension for 1 mi...

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Abstract

The invention discloses a molecular identification primer, kit and method for five groupers. The invention firstly provides the molecular identification primer for five groupers. The nucleotide sequence of the primer is as shown in SEQ ID NO: 1-2; and the five groupers are any one or more of Yunlong hybrid spot, tiger dragon hybrid spot, red dragon hybrid spot, epinephelus akaara or epinephelus coioides. By utilizing the primer, the five groupers can be quickly and effectively identified by adopting a method of combining mitochondrial genome DNA barcodes and a restriction endonuclease technology, and the primers are stable in reaction and good in repeatability; compared with the traditional morphological identification, the method has the characteristics of high detection accuracy and objectivity; compared with a simple mitochondrial genome DNA bar code sequencing identification method, the method has the characteristics of strong intuition and simple judgment; and therefore, the primer provided by the invention has a wide application prospect in identifying the five groupers or preparing a kit for identifying the five groupers.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology. More specifically, it relates to a molecular identification primer, kit and method for five species of grouper. Background technique [0002] Fish mitochondrial genomic DNA includes: 1 main non-coding region (D-loop), 1 light chain replication initiation region (O L ), 2 ribosomal RNA genes (12rRNA and 16S rRNA), 22 transfer RNA genes (tRNA) and 13 protein coding genes. Epinephelus spp. is the general name of Epinephelus, belonging to Osteichthyes, Perciformes, Serranidae, and Epinephelinate. There are more than 40 species of grouper recorded along the coast of my country. They are mainly produced in the South China Sea and the southern part of the East China Sea. Because of their delicious meat, rich nutrition, strong resistance to stress, and fast growth, they have become increasingly popular among consumers and farmers. Favor. In recent years, hybrid breeding has been widely used as an ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/158C12Q2565/125C12Q2563/185C12Q2521/301Y02A40/81
Inventor 杨敏王庆秦启伟王雨欣王黎
Owner SOUTH CHINA AGRI UNIV
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