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A primer for quickly identifying Cynoglossus semilaevis and Cynoglossus longirostris, a design method and an application method thereof

A semi-smooth tongue sole and application method technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve problems such as time-consuming and labor-consuming, species identification and system evolution confusion, and achieve cost savings, The effect of omitting the morphological identification procedure

Inactive Publication Date: 2019-01-11
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that there is currently no effective method for rapid and accurate identification of half-smooth tongue sole and long-nose red tongue sole, and only based on traditional morphological observation of their biomorphic characteristics, it is easy to cause confusion in species identification and phylogenetic evolution. The number of scales, the number of lateral line scales, and the number of vertebrae are time-consuming and labor-intensive to identify the differences. The present invention provides a primer for quickly identifying semi-smooth tongue sole and long-nose red-tongued sole and its design method and application method.

Method used

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  • A primer for quickly identifying Cynoglossus semilaevis and Cynoglossus longirostris, a design method and an application method thereof

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Embodiment

[0025] Preparation of primers:

[0026] S1) Download the rDNA sequences of half-smooth tongue sole and red tongue sole from the Genbank database;

[0027] S2) Perform multiple alignments on the downloaded rDNA sequences, and find relatively conserved sequences and regions with large variations at the 3' end and 5' end respectively;

[0028] S3) Design primers on the 18S rDNA and 28S rDNA conserved at both ends, and amplify two ITS1-5.8S-ITS2 sequences of tongue sole fishes with very similar shapes. The resulting primers are

[0029] Forward primer SEQ ID NO.1: 5'-TCGCTACTACCGATTGGATGGTTTA-3';

[0030] Reverse primer SEQ ID NO.2: 5'-GCTCTTCCCTCTTCACTCG-3'.

[0031] The primers prepared above were used to amplify and identify half-smooth tongue sole and red tongue sole, and the specific steps were as follows:

[0032] 1) Collection, identification and preservation of half-smooth tongue sole and red tongue sole

[0033] The samples of half-smooth tongue sole and long-nosed re...

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Abstract

The invention relates to the field of molecular biotechnology. In order to identify Cynoglossus semilaevis and Cynoglossus longirostris quickly and accurately, the present invention provides a primerfor quickly identifying Cynoglossus semilaevis and Cynoglossus longirostris, a design method and an application method thereof, which are only based on the traditional morphological observation and are easy to lead to problems such as species identification and phylogenetic confusion. The primers provided by the invention can efficiently and specifically amplify ITS1 of Cynoglossus semilaevis andCynoglossus longirostris. 5.8S-ITS2 rDNA sequence, which can amplify two species simultaneously at one time, has the advantages of high efficiency, rapidity and convenience, and the primers are used in ITS1-5.8S-After ITS2 rDNA sequence amplification, the length heterogeneity of ITS1 can be observed directly by gel electrophoresis, which saves the cost and improves the working efficiency and accuracy greatly.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a primer for quickly identifying half-smooth tongue sole and long-nose red tongue sole, a design method and an application method thereof. Background technique [0002] The half-smooth tongue sole and the long-nose red tongue sole belong to the genus Glossole sole of the family Pleuriformes. They both have a long tongue shape, two nostrils on the left side of the head, and three lateral lines on the left side of the body. They are very similar in shape. They are mainly distinguished by differences in the number of fin rays, lateral line scales, and vertebrae. However, because the scales are easy to fall off after leaving the water, it is difficult to identify the two based on traditional morphology, which leads to confusion in species identification and phylogenetic evolution; and by observing the differences in the number of fin rays, lateral line scales, and vertebrae...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124
Inventor 龚理孔晓瑜
Owner ZHEJIANG OCEAN UNIV
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