Method for analyzing phylogenetic development of babylonia formosae based on complete mitochondrial genomes

A mitochondrial genome and phylogeny technology, applied in the fields of genomics, biochemical equipment and methods, proteomics, etc., can solve problems such as confusion, misclassification, and vulnerability to environmental factors, and achieve convenient and effective analysis.

Pending Publication Date: 2020-01-21
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to overcome the phylogenetic relationship between the classification and the phylogenetic relationship of the Taiwan Dongfeng snail system of the Mollidae species by using morphological methods, because the engraving, color and shape of the shell surface are easily affected by environmental factors, and there is a relatively large problem. Instability, coupled with the convergent evolution of...

Method used

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  • Method for analyzing phylogenetic development of babylonia formosae based on complete mitochondrial genomes
  • Method for analyzing phylogenetic development of babylonia formosae based on complete mitochondrial genomes
  • Method for analyzing phylogenetic development of babylonia formosae based on complete mitochondrial genomes

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Embodiment 1

[0037] Embodiment 1: A method for analyzing the phylogeny of Taiwan Dongfeng snail based on the whole mitochondrial genome, comprising the following steps:

[0038] 1) Sample collection: Collect Taiwan Dongfeng snail, take 17mg Taiwan Dongfeng snail muscle tissue, add 450ul lysate, add 13ul Protein K, turn over, 57 ℃ water bath for 13h, then add 300ul NaCl solution with a concentration of 6M, vortex fully , placed at 4°C, centrifuged at 11,000 rcf, took the supernatant, added an equal volume of isopropanol, turned it over, and stored it in a freezer, then centrifuged and discarded the supernatant, dried the precipitate in an oven at 50°C after washing, and added 55ul of pure water Finally, the genomic DNA was detected by agarose gel electrophoresis; then the mitochondrial genomic DNA was extracted from the muscle tissue, frozen and stored at -25°C;

[0039] 2) Put the mitochondrial genomic DNA in an ultrasonic wave at 90W for 3s, repeat 4 times, and interrupt at an interval of...

Embodiment 2

[0044] Embodiment 2: A method for analyzing the phylogeny of Taiwan Dongfeng snail based on the whole mitochondrial genome, comprising the following steps:

[0045] 1) Sample collection: Collect Taiwan Dongfeng snail, take 15mg Taiwan Dongfeng snail muscle tissue, add 400ul lysate, add 10ul Protein K, turn over, 60 ℃ water bath for 12h, then add 300ul NaCl solution with a concentration of 6M, vortex fully , placed at 4°C, centrifuged at 10,000 rcf, took the supernatant, added an equal volume of isopropanol, turned it over, and stored it in a freezer, then centrifuged and discarded the supernatant, dried the precipitate in an oven at 45°C after washing, and added 50ul of pure water Finally, the genomic DNA was detected by agarose gel electrophoresis; then the mitochondrial genomic DNA was extracted from the muscle tissue and frozen at -30°C for cryopreservation;

[0046] 2) Put the mitochondrial genomic DNA in an ultrasonic wave under 80W for 2s, repeat 5 times, and interrupt a...

Embodiment 3

[0051] Embodiment 3: A method for analyzing the phylogeny of Taiwan Dongfeng snail based on the whole mitochondrial genome, comprising the following steps:

[0052] 1) Sample collection: Collect Taiwan Dongfeng snail, take 20mg Taiwan Dongfeng snail muscle tissue, add 500ul lysate, add 15ul Protein K, turn over, 55 ℃ water bath for 14h, then add 300ul NaCl solution with a concentration of 6M, vortex fully, Place at 4°C and centrifuge at 12000rcf, take the supernatant, add an equal volume of isopropanol, turn it over, and store it in a freezer, then centrifuge and discard the supernatant, wash the precipitate and dry it in an oven at 55°C, add 60ul of pure water , using agarose gel electrophoresis to detect genomic DNA; then extract mitochondrial genomic DNA from muscle tissue, freeze and store at -15°C;

[0053] 2) Put the mitochondrial genomic DNA in an ultrasonic wave under 100W for 4s, repeat 3 times, and interrupt at an interval of 10s, after purification, perform DNA end ...

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Abstract

The invention discloses a method for analyzing phylogenetic development of babylonia formosae based on complete mitochondrial genomes, and relates to the field of analysis of the phylogenetic development of the babylonia formosae. The method for analyzing phylogenetic development of the babylonia formosae based on the complete mitochondrial genomes comprises the following steps: 1) sample collection: collecting babylonia formosae extracting mitochondrial genomes DNA; 2), disrupting and purifying the mitochondrial genomes DNA and then building a DNA sequencing library; 3), obtaining a DNA cluster; 4), sequencing the babylonia formosae mitochondrial genomes DNA; 5), comparing the babylonia formosae mitochondrial genomes with other babylonias having known sequences, and then noting encoded protein genes; and 6), comparing a nucleotide sequence of each gene of the babylonia formosae extracting mitochondrial genomes DNA and an amino acid sequence deduced by the nucleotide sequence with other caenogastropoda species, and performing series connection and then analyzing the phylogenetic development. According to the phylogenetic analysis method of the babylonia formosae based on the complete mitochondrial genomes disclosed by the invention, based on the complete mitochondrial genomes, the phylogenetic development of the babylonia formosae is analyzed, complete data of the complete mitochondrial genomes of the babylonia formosae are obtained, and thus, phylogenetic development is performed, and analysis is performed conveniently and effectively.

Description

technical field [0001] The invention relates to the field of phylogenetic analysis of Taiwan Dongfeng snails, in particular to a method for analyzing the phylogeny of Taiwan Dongfeng snails based on the whole mitochondrial genome. Background technique [0002] Taiwan Dongfeng snail (Babylonia formosae) belongs to the mollusc phylum, Gastropoda, Mollidae, Dongfeng snail genus. The taxonomy and phylogenetic relationship of Mollidae mainly depend on the morphological characteristics of shells and shells so far. However, because the engraving, color and shape of the shell surface are easily affected by environmental factors, there is greater instability, coupled with the convergent evolution of similar species, the shells of the Mollidae species tend to be simplified, so they are not Do not provide enough distinguishing features, so classification based solely on the shell and shell morphology often leads to errors and confusion in classification. The phylogenetic relationship...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12Q1/6806G16B20/00
CPCC12Q1/6869C12Q1/6806G16B20/00C12Q2535/122C12Q2563/107C12Q2523/301C12Q2525/191C12Q2531/113
Inventor 叶莹莹许中天冯鉴童
Owner ZHEJIANG OCEAN UNIV
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