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Amplification primer and amplification method of complete mitochondrial genome sequence of nibea japonic

A whole genome, amplification primer technology, applied in the field of genetic engineering

Active Publication Date: 2015-12-16
MARINE FISHERIES RES INST OF ZHEJIANG
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Problems solved by technology

So far, there are no reports on the mitochondrial genome sequence, genetic and germplasm resources identification and conservation of Japanese yellowtail

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0039] Using the extracted Japanese croaker genomic DNA as a template, first use 4 pairs of specific primers (pairs 1, 6, 11 and 16) to amplify 4 large fragment products, and then use the method of genome walking, using the method of the present invention (2-5, 7-10, 12-15, 17-20) primers were used for PCR amplification, and the PCR products were detected by 1.0% agarose electrophoresis and then sent to the biological company for sequencing. The sequencing results were spliced ​​using biological software. Specific steps are as follows:

[0040] 1. 5×TBEBuffer preparation

[0041] Weigh tromethamine 54g, Na 2 EDTA·2H 2 O3.72g, boric acid 27.5g in a 1L beaker; add about 800ml deionized water, stir evenly; adjust pH to 8.3 with NaOH, add deionized water to 1L, store at room temperature for later use.

[0042] 2. DNA extraction

[0043] Using the high-salt method to extract the DNA of Japanese croaker, the steps are as follows:

[0044] (1) Take 50 mg of the back muscle tissu...

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PUM

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Abstract

The invention discloses an amplification primer and an amplification method of complete mitochondrial genome sequence of nibea japonic; through PCR amplification on the genome of nibea japonic by virtue of 20 pairs of specific primers, the complete mitochondrial genome sequence of the nibea japonic can be obtained, so that material basis is provided for conservation genetics and evolutionary genetics of nibea japonic as well as for molecular marker-assisted selection of excellent breeding varieties.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to an amplification primer and an amplification method for the whole mitochondrial genome sequence of Japanese yellow croaker. Background technique [0002] Japanese croaker ( Nibeajaponic ) commonly known as black cod, Perciformes (Perciformes), Sciaenidae (Sciaenidae), yellow croaker ( Nibea ) (Zhu Yuanding, 1963; Cheng Qingtai, 1987), is a large and high-quality edible fish, mainly distributed in the East China Sea, South China Sea and the southern coast of Japan. [0003] Japanese yellow croaker is a large edible fish. The largest individual can reach more than 1.5 meters. It can adapt to adverse environments and resist diseases. It is easy to breed, has a low bait coefficient, and grows rapidly. One of the main objects. Due to its high economic value, the fishing pressure on Japanese croaker has continued unabated. In recent years, due to the decline of resourc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/16
Inventor 胡则辉柴学军王跃斌朱云海
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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