Amplification primer and amplification method of complete mitochondrial genome sequence of nibea miichthioides chu

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

Inactive Publication Date: 2017-04-26
MARINE FISHERIES RES INST OF ZHEJIANG
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  • Abstract
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  • Application Information

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Problems solved by technology

So far, there has been no report on the mitochondrial genom

Method used

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Experimental program
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Effect test

Embodiment

[0042] Using the extracted genomic DNA of yellow aquarium as a template, first amplify the products of 4 large fragments with 4 pairs of specific primers for large fragment amplification, and then use the method of genome walking to use the amplified 4 large fragments respectively. The fragment was used as a template for PCR amplification using 19 primers for genome walking. The PCR product was detected by 1.0% agarose electrophoresis and sent to the biological company for sequencing. The sequencing results were assembled using biological software. The specific steps are as follows:

[0043] 1. 5×TBE Buffer preparation

[0044] Weigh 54g of tromethamine, Na 2 EDTA·2H 2 O 3.72g, 27.5g boric acid in a 1L beaker; add about 800ml deionized water and stir evenly; adjust the pH to 8.3 with NaOH, add deionized water to a constant volume to 1L, and store at room temperature for later use.

[0045] 2. DNA extraction

[0046] The high-salt method is used to extract DNA from koala croaker, the s...

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PUM

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Abstract

The invention discloses an amplification primer and an amplification method of a complete mitochondrial genome sequence of nibea miichthioides chu. The amplification primer comprises 4 klenow fragment amplification primer pairs and 19 genome walking primers. According to the amplification primer and the amplification method provided by the invention, the complete mitochondrial genome sequence of nibea miichthioides chu, which is obtained by conducting PCR amplification on genome of the nibea miichthioides chu, can provide a material base for conservation genetics and evolutionary genetics of the nibea miichthioides chu and for marker-assisted selection of high-quality breeding varieties.

Description

Technical field [0001] The present invention relates to the technical field of genetic engineering, in particular to an amplification primer for the whole mitochondrial genome sequence of croaker croaker and its amplification method. Background technique [0002] Nibea miichthioides Chu (Lo&Wu), also known as croaker and perch, belongs to the genus Perciformes, Tothocephalidae, and Yellow croaker. It is mainly distributed in the south-central East my country Sea and the South China Sea. It is a coastal warm Large carnivorous fish in the middle and lower waters. Because of its strong disease resistance and fast growth, it has become an excellent cage culture species along the coasts of Fujian, Zhejiang and Guangdong. [0003] Owl-shaped yellow croaker is a large edible fish, the largest individual can reach 1.5 meters. Due to long-term indiscriminate fishing and habitat destruction, the resources of Ophiopogon japonicus are declining. The artificially cultivated Ophiopogon japonicu...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10
CPCC12N15/11C12N15/10C12Q2531/113
Inventor 胡则辉柴学军王跃斌胡成硕
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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