Amplimer of large yellow croaker mitochondrion complete genome sequence and application thereof

A whole genome and sequence amplification technology, which is applied in the field of amplification primers for the whole mitochondrial genome sequence of large yellow croaker, can solve the problems of unmeasured and achieve high success rate and convenient splicing

Active Publication Date: 2012-11-14
NINGBO UNIV
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  • Application Information

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Problems solved by technology

The Chinese patent name is the complete mitochondrial genome sequence and its amplification primers of the salmon salmon (application number CN200910073114.8), and the cloning and evolution analysis of the complete mitochondrial genome sequence of wild boar in Heilongjiang Provinc...

Method used

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  • Amplimer of large yellow croaker mitochondrion complete genome sequence and application thereof
  • Amplimer of large yellow croaker mitochondrion complete genome sequence and application thereof
  • Amplimer of large yellow croaker mitochondrion complete genome sequence and application thereof

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specific Embodiment 1

[0032] The amplification primers of the large yellow croaker mitochondrial whole genome sequence of the present invention are 15 pairs, and the gene sequence is as follows:

[0033] The amplification product of the primers designed in the present invention should cover the entire mitochondrial genome, the length of the amplification product of each pair of primers is 900-1560 bp, and the sequence overlap obtained by two adjacent pairs of primers is more than 80 bp.

[0034]

specific Embodiment 2

[0035] Design method of amplification primers for whole mitochondrial genome sequence of large yellow croaker

[0036] 1. Screening of templates

[0037] Log in to the BCBI (http: / / www.ncbi.nlm.nih.gov / ) website to search for the complete mitochondrial genome sequence of the species belonging to the same family or even the same genus as the species to be tested. If multiple sequences are available, use blast The software compares and preferentially selects the sequence of the species closest to the species to be tested.

[0038]2. Design primers according to the template sequence

[0039] Use the screened sequence as a template, and use the PCR primer design software Primer Premier5.0 to design primers for PCR amplification. The specific ideas are as follows:

[0040] 1) The length of PCR amplification should be less than 1600bp, because the current mainstream first-generation sequencer ABI3730 sequencing read length can reach 800 to 1000bp, and the sequencing accuracy...

specific Embodiment 3

[0045] The present invention utilizes 15 pairs of amplification primers for the whole mitochondrial genome sequence of large yellow croaker to obtain the whole mitochondrial genome sequence of Daiqu family large yellow croaker, as shown in SEQ ID NO:31.

[0046] The whole mitochondrial gene sequence of Daiqu large yellow croaker is 16467bp in total, including 22 tRNAs, 2 rRNAs, 13 protein-coding genes and 1 D-loop region.

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Abstract

The invention discloses an amplimer of a large yellow croaker mitochondrion complete genome sequence and an application thereof. The 15 pairs of amplimer are applied, a gene order is respectively shown as SEQ ID No: 1-30, the amplimer is further used for obtaining a Tai Qu family large yellow croaker mitochondrion complete genome sequence and a Min-Yue East family large yellow croaker mitochondrion complete genome sequence which are respectively shown as SEQ ID No: 31 and SEQ ID No: 32, and an amplimer of large yellow croakers in a high variation region is obtained. The amplimer and the application have the advantages that the Tai Qu family large yellow croaker and Min-Yue East family large yellow croaker mitochondrion complete genome sequences can be quickly and efficiently obtained respectively through the 15 pairs of amplimers; and the amplimer of the large yellow croakers in the high variation region provides support for the research of species identification, geographical population identification, germ plasm resources and genetic diversity of the large yellow croakers, and further provide the basis for developing an identification technology of Tai Qu family large yellow croakers and Min-Yue East family large yellow croakers.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer for amplifying the whole mitochondrial genome sequence of large yellow croaker and an application thereof. Background technique [0002] Large yellow croaker (Pseudosciaena crocea), also known as yellow croaker and yellow croaker, belongs to Perciformes (Perciformes), Sciaenidae (Sciaenidae), and Pseudosciaena (Pseudosciaena). It is the most productive fish species in marine cage culture. The coastal large yellow croaker in my country can be roughly divided into three geographical groups: Daiqu, Fujian-Yuedong and Naozhou. The large yellow croaker of the Daiqu family is golden or tiger yellow, shiny, with bright red or purple gills, full eyeballs, strong muscles, and high elasticity. , due to overfishing and environmental degradation, the wild Daiqu large yellow croaker is very scarce. Due to the similar external shape of the large yellow croaker of the Fujian-Yuedong fam...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 沈锡权闫旭红王梦林严小军
Owner NINGBO UNIV
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