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A Universal Primer and Its Design and Amplification Method for Transcriptional Spacer 2 in Ribosome Ribosome of Pleuriformes Soleidae Fish

An internal transcribed spacer, universal primer technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. The effect of obvious difference, efficient and quick identification

Active Publication Date: 2021-06-18
ZHEJIANG OCEAN UNIV
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Problems solved by technology

[0004] On September 29, 2017, the Chinese Patent Office disclosed the invention authorization of a spider mitochondrial genome full-sequence amplification primer and amplification method, the authorization publication number: CN104531688B. Long PCR (L-PCR) amplification and sequencing can identify spiders, but it can only amplify spiders; it also uses DNA technology to identify organisms, but it cannot identify specific biological species, and must Only by sequencing can it be accurately determined, and it is necessary to design multiple pairs of primers to amplify the entire genome sequence, which takes a long time and is cumbersome.

Method used

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  • A Universal Primer and Its Design and Amplification Method for Transcriptional Spacer 2 in Ribosome Ribosome of Pleuriformes Soleidae Fish

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Embodiment Construction

[0024] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention and the accompanying drawings. Apparently, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0025] (1) Collection, identification and preservation of fish samples of Pleuriformes Soleidae

[0026] All the fish samples of Pleuriformes Soleidae used in the examples were collected from the natural environment in the wild, and all the collected specimens were brought back to the laboratory for information registration, and all were identified with a stereo microscope to determine the species. The samples of Plaiceiformes soles that had been...

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Abstract

The present invention relates to a kind of general primer and design and amplification method for transcriptional spacer 2 in the ribosome ribosome of Plaiceidae fish which can be used for rapid identification of Plaiceidae fishes, and the provided ribose ribose The in vivo transcriptional spacer 2 universal primer can efficiently and specifically amplify the ITS2 of the flounder sole family Octopus octopus, and can simultaneously amplify multiple species at one time, which has the advantages of high efficiency, rapidity and convenience, and because ITS2 The sequence lengths of different species of Soleidae are significantly different, and can be directly observed with the naked eye by gel electrophoresis technology, so as to efficiently and quickly identify species with similar or morphological damage.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a general primer for transcriptional spacer 2 in ribosome ribosomes and a method for designing and amplifying them, which can be used for rapid identification of fishes of the family Soleidae. Background technique [0002] Soleidae, as the most abundant group of flatfishes, has about 35 genera and 175 species. Many species have very important economic value. The study of the transcriptional spacer 2 in the ribosome is even more blank. [0003] The internal transcribed spacer 1 (ITS2) is a non-coding region located between the 5.8S rDNA and 28S rDNA of the ribosomal RNA coding gene. The evolution rate of 5.8S rDNA and 28S rDNA is slow, and the sequence is very conservative, while the evolution rate of ITS2 is relatively fast, basically tends to be similar within the species, but shows significant differences between species, so it can be used as a tool to distinguish di...

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Application Information

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