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Amplification Primers and Amplification Method of the Whole Mitochondrial Genome Sequence of Japanese Yellow Mackerel

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

Active Publication Date: 2018-06-15
MARINE FISHERIES RES INST OF ZHEJIANG
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

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×TBE Buffer preparation

[0041] Weigh tromethamine 54g, Na 2 EDTA·2H 2 Put 3.72g of O and 27.5g of boric acid in a 1L beaker; add about 800ml of deionized water and stir evenly; adjust the pH to 8.3 with NaOH, add deionized water to make the volume to 1L, and 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: ...

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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 ( Nibea japonic ) 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 resour...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
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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