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Simple efficient method for extracting oncomelania mitochondrial genome DNA

An extraction method and genome technology, applied in the field of extraction of Oncomelania mitochondrial genomic DNA, can solve the problems of human poisoning, nuclear DNA pollution, complicated operation process, etc., and achieve the effect of improving yield and simple process

Active Publication Date: 2016-12-21
WANNAN MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] When extracting Oncomelania mitochondrial genomic DNA, commonly used extraction methods include sucrose density gradient centrifugation, alkali lysis, Triton method, high-salt precipitation method and kits, etc. These extraction methods have complex operation process, low yield, and harm to the human body. Poisonous, serious nuclear DNA contamination, expensive reagents and other disadvantages

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  • Simple efficient method for extracting oncomelania mitochondrial genome DNA
  • Simple efficient method for extracting oncomelania mitochondrial genome DNA
  • Simple efficient method for extracting oncomelania mitochondrial genome DNA

Examples

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Effect test

Embodiment 1

[0035] A simple and efficient method for extracting mitochondrial genomic DNA of Oncomelania snails, comprising the following steps:

[0036] a) Grinding process:

[0037] Remove the shell and digestive system tissue from a single snail, take 2 mg of gastropod muscle tissue, put it into a grinder, add 50 μl of homogenate, and grind the gastropod muscle tissue up and down for 1-2 minutes to obtain a homogenate containing muscle tissue. Transfer the homogenate containing muscle tissue to a centrifuge tube, then add 50 μl of homogenate, grind the gastropod muscle tissue up and down, repeat 3 times, grind until the gastropod muscle becomes 5-30 μm particles, and finally wash with 100 μl of homogenate A grinder to form a flushing liquid, combining the homogenate containing muscle tissue obtained by grinding several times with the flushing liquid to form a ground homogenate;

[0038] b) cracking process:

[0039] Put 300 μl of ground homogenate into a 400W microwave oven, select h...

Embodiment 2

[0046] A simple and efficient method for extracting mitochondrial genomic DNA of Oncomelania snails, comprising the following steps:

[0047] a) Grinding process:

[0048]Remove the shell and digestive system tissue from a single snail, take 2.5 mg of gastropod muscle tissue, put it into a grinder, add 80 μl of homogenate, grind the gastropod muscle tissue up and down for 1-2 minutes, and obtain a homogenate containing muscle tissue , transfer the homogenate containing muscle tissue to a centrifuge tube, then add 80 μl of homogenate, grind the gastropod muscle tissue up and down, repeat 2 times, grind until the gastropod muscle becomes 5-30 μm particles, and finally use 100 μl of homogenate Rinse the grinder to form a rinsing liquid, and combine the homogenate containing muscle tissue obtained by grinding several times with the rinsing liquid to form a ground homogenate;

[0049] b) cracking process:

[0050] Put 340 μl of ground homogenate into a 400W microwave oven, select...

Embodiment 3

[0057] A simple and efficient method for extracting mitochondrial genomic DNA of Oncomelania snails, comprising the following steps:

[0058] a) Grinding process:

[0059] Remove the shell and digestive system tissue from a single snail, take 3 mg of the gastropod muscle tissue, put it into a grinder, add 100 μl of homogenate, and grind the gastropod muscle tissue up and down for 1-2 minutes to obtain a homogenate containing muscle tissue. Transfer the homogenate containing muscle tissue to a centrifuge tube, then add 100 μl of homogenate, grind the gastropod muscle tissue up and down, repeat twice, grind until the gastropod muscle becomes 5-30 μm particles, and finally wash with 100 μl of homogenate A grinder to form a flushing liquid, combining the homogenate containing muscle tissue obtained by grinding several times with the flushing liquid to form a ground homogenate;

[0060] b) cracking process:

[0061] Put 400 μl of ground homogenate into a 400W microwave oven, sele...

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Abstract

The invention discloses a simple efficient method for extracting an oncomelania mitochondrial genome DNA. The method comprises the following steps: a) grinding process; b) splitting decomposition process; c) precipitation process; and d) purification process. Compared with the prior art, the yield is increased through the steps of pyrolysis, varied-temperature digestion of protease K, and precipitation and purification of potassium acetate. The quality of the extracted mitochondrial genome DNA is detected through an ultraviolet spectrophotometer, agarose gel electrophoresis and PCR (polymerase chain reaction) detection. The result shows that the concentration and the purity of the extracted mitochondrial genome DNA both meet the requirements of researches on population genetic structure, system evolution, geographical distribution and construction of a gene library. The establishment of the method leads to simpler and more efficient process of extracting the mitochondrial genome DNA of small shellfish like an oncomelania.

Description

technical field [0001] The invention belongs to a method for extracting parasite DNA, in particular to a method for extracting mitochondrial genome DNA of oncomelania. Background technique [0002] As the only intermediate host of Schistosoma japonicum, the distribution of oncomelania is easily affected by various factors such as water quality, water temperature, and climate, resulting in geographical isolation, resulting in multiple species in different geographical groups or even in sympatrically distributed groups. Therefore, the current status of genetic polymorphism within and between populations of Oncomelania snails, the intensity of gene flow between populations, and the genetic variation and differentiation among populations have become the focus of the prevention and control of schistosomiasis. Compared with the nuclear genome, the mitochondrial genome has its unique genetic characteristics, such as maternal inheritance, lack of recombination, and fast evolution ra...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q2521/537C12Q2527/125
Inventor 孙恩涛王康许树俊王忆楠
Owner WANNAN MEDICAL COLLEGE
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