Whole genome DNA of scylla paramamosain mitochondria and testing method

A mitochondrial gene, whole genome technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of low genetic information and misleading research results, and achieve the effect of low cost and easy to master

Inactive Publication Date: 2013-04-24
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

However, the genetic information revealed by single-gene markers is much lower than that provided by whole genomes and has the potential to mislead research results

Method used

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  • Whole genome DNA of scylla paramamosain mitochondria and testing method
  • Whole genome DNA of scylla paramamosain mitochondria and testing method
  • Whole genome DNA of scylla paramamosain mitochondria and testing method

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Embodiment 1

[0023] 1. Collection and analysis of mitochondrial-related gene sequences of Scylla pseudocavena

[0024] First, on the one hand, the present invention screens the mitochondrial-related gene sequences of Scylla pseudocarpus from the transcriptome library obtained by sequencing; The mitochondrial genome sequences of closely related species Scylla serrata and Portunus trituberculatus were compared, and the relative positions of related genes in the mitochondrial genome sequences of closely related species were analyzed.

[0025] 2. Primer design and PCR amplification

[0026] According to the relative position of the mitochondrial gene in the whole genome sequence of the closely related species, the primers were designed using the Scylla pseudosatis gene sequence as a template, and the unknown sequence between two adjacent genes was amplified by PCR method. The PCR amplification reaction system is 25 μl, including 1 μl of genomic DNA template, upstream and downstream primers wi...

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Abstract

The invention relates to a whole genome DNA of scylla paramamosain mitochondria and a testing method. The DNA molecule which is in a closed ring structure has a length of 15824 bp; and the testing method comprises the steps of: (1) collecting relative gene sequences of the scylla paramamosain mitochondria and comparing the relative gene sequences of the scylla paramamosain mitochondria with whole genome sequences of mitochondria of sibling species; (2) designing a primer and carrying out PCR (Polymerase Chain Reaction) amplification; and (3) sequencing PCR products and assembling the sequences. According to the invention, the whole genome DNA sequences of the scylla paramamosain mitochondria are obtained. The testing method disclosed by the invention has the advantages of being fast in speed, low in cost, easy to master and the like; and the obtained whole genome DNA of the mitochondria can be applied to the fields such as heritable variation analysis of the scylla paramamosain, genetic resource authorization and protection, phylogeny and the like.

Description

technical field [0001] The invention belongs to the field of Scylla pseudomae genomics, and in particular relates to a whole genome DNA of the Scylla pseudomae mitochondria and a detection method. Background technique [0002] In most multicellular animals, mitochondrial genomic DNA is a typical closed circular structure with a length between 14 and 18 kb, capable of autonomous replication and transcription. The mitochondrial genome usually encodes 37 genes, including: 13 protein-coding genes, 22 tRNA genes, and 2 rRNA genes. In addition, it also contains a large non-coding region (control region). The whole mitochondrial genome can not only provide more genetic information than a single gene, but also reveal the characteristics of the genome level, which helps to better understand the characteristics of genome evolution and development. So far, scholars have revealed the whole mitochondrial genome DNA information of many crustaceans, including: Penaeus monodon, Penaeus chi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12Q1/68
Inventor 马洪雨马凌波马春艳
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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