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Specific primer sequence capable of being applied in method for identifying different fish, and DNA (Deoxyribose Nucleic Acid) molecular marker method for identifying different fish

A technology of DNA molecules, labeling methods, used in recombinant DNA technology, DNA/RNA fragments, biochemical equipment and methods, etc.

Active Publication Date: 2014-02-19
HUNAN NORMAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the genetic diversity of the 5S rDNA gene transcriptional spacer region has been used for molecular evolution and phylogenetic research in many applications in plant research, but 5S has not been used Genetic diversity of rDNA gene transcribed spacer region for rapid identification of different fish species reported

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  • Specific primer sequence capable of being applied in method for identifying different fish, and DNA (Deoxyribose Nucleic Acid) molecular marker method for identifying different fish
  • Specific primer sequence capable of being applied in method for identifying different fish, and DNA (Deoxyribose Nucleic Acid) molecular marker method for identifying different fish
  • Specific primer sequence capable of being applied in method for identifying different fish, and DNA (Deoxyribose Nucleic Acid) molecular marker method for identifying different fish

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Embodiment

[0029] Referring to the known coding sequence of the 5S rDNA gene of other fish, a specific primer sequence that can be used in the DNA molecular marker method for differentiating fish was designed with Primer Premier 5.0 software and Jellyfish 1.4 software, provided by Shanghai Sangong Co., Ltd. Company (Shanghai Sangon for short) synthesized, the specific primer sequence is as follows:

[0030] Upstream primer: 5'-GCCCGATCTCGTCTGATCTCG-3';

[0031] Downstream primer: 5'-GCGCTCAGGTTGGTATGGCCG-3'.

[0032] A kind of DNA molecular marker method of different fishes with above-mentioned specific primer sequence identification of the present invention, comprises the following steps:

[0033] 1. Prepare experimental materials.

[0034] Angelfish ( Pterophyllum ),carp( Cyprinus carpio L. ), yellow-tailed pomfret ( Xenocypris davidi Bleeker ), Crawfish ( Erythroculter ilishaeformis Bleeker ), moray eel ( Muraenesox cinereus ), Japanese eel ( Anguilla Japonica ), mandarin ...

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Abstract

The invention discloses a specific primer sequence capable of being applied in a DNA (Deoxyribose Nucleic Acid) molecular marker method for identifying different fish. The specific primer sequence comprises an upstream primer and a downstream primer. The DNA molecular marker method for identifying different fish by using the specific primer sequence comprises the steps that a specific primer is designed according to a 5S rDNA genic conservative coding region sequence of the given fish; a genome of a fish material to be identified is extracted; a PCR (Polymerase Chain Reaction) system is designed by taking DNA of the genome as a template; a 5S rDNA gene of the system is amplified; a DNA segment mode of the sample 5S rDNA gene is obtained after PCR amplified reaction, agarose gel electrophoresis and DNA gel recovery kit purification by utilizing the polymorphyism of a nontranscribed spacer of the 5S rDNA gene; and finally the kind of the fish material to be identified is judged by comparing the DNA segment mode in gel imaging with the given fish. The method has the advantages that the method is rapid, accurate, and easy and simple to operate.

Description

technical field [0001] The invention relates to a method for analyzing genetic characteristics and molecular genetic relationship of fish by using molecular genetic markers, in particular to a method for identifying and analyzing different fishes by using DNA molecular markers based on 5S rDNA gene DNA band patterns combined with DNA sequencing. Background technique [0002] Compared with morphological and cell biological methods, molecular biology techniques have advantages in identifying and analyzing the genetic characteristics of individuals or populations. Biologists have long sought a faster, more accurate, and more stable molecular biology method for labeling and identifying different animals. Although whole-genome sequencing can provide valuable genetic information, it is unrealistic to sequence all species in the biological kingdom. Therefore, molecular genetic markers are still the main means of studying genetic diversity and its variation at the individual or pop...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 刘少军覃钦博刘筠王余德王娟
Owner HUNAN NORMAL UNIVERSITY
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