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The specific primer sequence that can be applied in the method for identifying different fish and the dna molecular marker method for identifying different fish

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

Active Publication Date: 2016-04-13
HUNAN NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, molecular evolution and phylogenetic studies using the genetic diversity of the 5SrDNA gene transcriptional spacer have been widely used in plant research, but there has been no report on the use of the genetic diversity of the 5SrDNA gene transcriptional spacer for rapid identification of different fishes

Method used

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  • The specific primer sequence that can be applied in the method for identifying different fish and the dna molecular marker method for identifying different fish
  • The specific primer sequence that can be applied in the method for identifying different fish and the dna molecular marker method for identifying different fish
  • The specific primer sequence that can be applied in the method for identifying different fish and the dna molecular marker method for identifying different fish

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Embodiment

[0029] Referring to the known coding region sequences of 5S rDNA genes of other fishes, a specific primer sequence that can be used in the DNA molecular marker method for differentiating different fishes was designed with PrimerPremier5.0 software and Jellyfish1.4 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 ( Erythroculterilisha eformis Bleeker ), moray eel ( Muraenes oxcinereus ), Japanese eel ( Anguilla Japo...

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Abstract

The invention discloses specific primer sequences that can be used in DNA molecular marker methods for differentiating different fish, including upstream primers and downstream primers. Using the specific primer sequences to identify different fish DNA molecular marker methods includes: first according to known 5S of fish? Design specific primers for the conserved coding region sequence of the rDNA gene; then extract the genome of the fish material to be identified; use the genomic DNA as a template to design a PCR reaction system to amplify its 5S? rDNA gene, using 5S? After the polymorphism of the non-transcribed spacer region of the rDNA gene was purified by PCR amplification reaction, agarose gel electrophoresis and DNA gel recovery kit, the sample 5S? The DNA fragment pattern of the rDNA gene; finally, the species of the fish material to be identified was determined by comparing the DNA fragment pattern in the gel imaging with the known fish. The method of the invention has the advantages of quickness, accuracy, easy operation and the like.

Description

technical field [0001] The invention relates to a method for using molecular genetic markers to analyze genetic characteristics and molecular genetic relationships of fish, in particular to a method for identifying and analyzing different fishes by using DNA molecular markers based on 5SrDNA 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 popul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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