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Whole genome sequencing method for mitochondria of Bagariusyarrelli Sykes

A technology of whole genome sequencing and giant fish, which is applied in the fields of biochemical equipment and methods, microbial determination/inspection, etc.

Active Publication Date: 2015-06-24
HONGHE COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the whole mitochondrial gene cloning technology at home and abroad.

Method used

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  • Whole genome sequencing method for mitochondria of Bagariusyarrelli Sykes
  • Whole genome sequencing method for mitochondria of Bagariusyarrelli Sykes
  • Whole genome sequencing method for mitochondria of Bagariusyarrelli Sykes

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Embodiment 1

[0022] A method for sequencing the whole mitochondrial genome of giant mandarin fish, comprising the following steps:

[0023] (1) Extraction of genomic DNA of giant chondrodfish: Take 0.1g-0.5g of fin ray tissue into a 1.5ml centrifuge tube, add 600μl of DNA extraction solution, then add proteinase K8-10μl and 5μl RNase, and gently rotate up and down to mix ; Then add an equal volume of phenol / chloroform / isoamyl alcohol with a volume ratio of 25:24:1, gently rotate up and down to mix well, centrifuge at 12000 rpm for 10 minutes at 4°C, take the supernatant, and then add an equal volume The volume ratio of chloroform / isoamyl alcohol is 24:1, gently rotate up and down to mix, and centrifuge at 12,000 rpm at 4°C for 10 minutes; add 2 times the volume of ice-cold absolute ethanol to the supernatant to precipitate DNA for 10 -25 minutes, rinse the pellet with 70% ethanol, dissolve the DNA with TE, and obtain the giant mandrill genome DNA, such as figure 1 shown; stored at -20°C f...

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Abstract

The invention discloses a whole genome sequencing method for mitochondria of Bagariusyarrelli Sykes, belonging to the technical field of aquatic organisms. The whole genome sequencing method comprises the following steps: extracting DNA of genomes of Bagariusyarrelli Sykes, and designing 18 pairs of homologous primers; carrying out PCR amplification, recycling target fragments, and carrying out carrier conjugation, conversion, cloning and sequencing; and carrying out sequence splicing to obtain overall-length mitochondrion gene sequence of Bagariusyarrelli Sykes. According to the whole genome sequencing method provided by the invention, the cloning of overall-length mitochondrion genes of Bagariusyarrelli Sykes is facilitated, and convenience is provided for the analysis of population genetics; the whole genome sequencing method is suitable for evaluating the molecular genetics and the group structure of the mitochondrion genes in Bagariusyarrelli Sykes; foundational data is provided for the resource protection of Bagariusyarrelli Sykes.

Description

technical field [0001] The invention belongs to the technical field of aquatic organisms, and in particular relates to a method for sequencing the whole mitochondrial genome of the giant carp fish. Background technique [0002] The giant chirp (Bagarius yarrelli Sykes) belongs to the Siluriformes, Bagarius family, and the genus Bagarius. Commonly known as "yellow croaker", the flesh is yellow, the individual is very large, and its feeding habits are mainly shrimp, loach, small fish and aquatic insects. Through preliminary studies on its external morphology, digestive system, reproductive and living habits, etc., it has been determined that the giant chinchilla is a bottom-dwelling carnivorous freshwater fish, and its digestive system and external morphological and structural characteristics have good adaptability. Mandrills produce buoyant eggs and have good reproductive characteristics. In the Lancang River, Nujiang River, and Yuanjiang River in Yunnan Province, there are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 杜民刘艳红周传江牛宝珍李娜
Owner HONGHE COLLEGE