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Method for harmlessly extracting Chinese sturgeon DNA

A Chinese sturgeon, non-damaged technology, applied in the field of bioengineering, can solve the problem of non-damaged DNA extraction from Chinese sturgeon, which is not reported, and achieve the effects of stable and reliable results, low cost and high quality

Active Publication Date: 2016-03-16
CHINESE STURGEON RES INST CHINA THREE GOR
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are only a few reports on the non-destructive extraction of DNA from fish, and there is no report on the non-destructive extraction of DNA from Chinese sturgeon

Method used

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  • Method for harmlessly extracting Chinese sturgeon DNA
  • Method for harmlessly extracting Chinese sturgeon DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A method for extracting Chinese sturgeon DNA without damage, said method comprising the steps of:

[0032] 1) Obtaining samples: Take 6 Chinese sturgeons from the breeding pond of the Chinese Sturgeon Research Institute in Yichang, Hubei, pick them up from the water, and gently scrape the surface of the Chinese sturgeon with your fingers. The direction of scraping is from head to tail. Put the mucus scraped on your fingers into a centrifuge tube, and after taking 200ul, put the centrifuge tube containing the mucus at 0°C to prevent DNA degradation;

[0033] 2) Sample processing: centrifuge the mucus in step 1), remove the supernatant, and obtain the centrifuged mucus;

[0034] 3) Cell lysing: add the lysate and proteinase K to the centrifuged mucus in step 2), and bathe in water at 56°C for 1 hour to rapidly lyse the cells to obtain the mucus after lysing the cells;

[0035] 4) Precipitate protein: put saturated phenol into the mucus after the lysed cells in the above ...

Embodiment 2

[0043] The detection of embodiment 2DNA

[0044] 1. Electrophoresis detection: Take 5ul of the extracted DNA and test it in 1% agarose gel electrophoresis, 120V voltage for 20 minutes, and observe whether there is a DNA band near the sample point through the gel imaging analysis system, the result is as follows figure 1 As shown, there is a white DNA band near the sample point, and the DNA band is clear without tailing, indicating that the DNA extraction is better.

[0045] 2. PCR detection: use the extracted DNA as a template to detect with microsatellite primers. The PCR amplification system is: 12.5ul of PCR reaction solution, 1ul of upstream and downstream primers, 9.5ul of ultrapure water, and 1ul of DNA template. The PCR reaction program was: pre-denaturation at 95°C for 2 min; denaturation at 95°C for 30 s, annealing temperature for 30 s, extension at 72°C for 30 s, 35 cycles; extension at 72°C for 10 min; storage at 4°C. Put the PCR amplification product into 1% a...

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Abstract

The invention relates to the field of bioengineering, in particular to a method for harmlessly extracting Chinese sturgeon DNA. The method includes the steps of obtaining a sample, processing the sample, conducting cell lysis, precipitating proteins, precipitating DNA, washing DNA and obtaining DNA. The complete method for extracting Chinese sturgeon mucus is established, Chinese sturgeon DNA can be rapidly and effectively extracted, extracted DNA stripes are clear and free of trailing, and it is shown that extracted DNA is high in quality and free of protein contamination and stable and reliable in result.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for extracting Chinese sturgeon DNA without damage. Background technique [0002] Chinese sturgeon Acipensersinensis It is an ancient large-scale migratory fish in the Haihe River. It was once distributed in the Yangtze River and Pearl River rivers in my country. Due to overfishing, environmental pollution, water conservancy project construction and other human reasons in recent years, the number of Chinese sturgeon has declined sharply. Now the Pearl River Basin The Chinese sturgeon has become extinct, and the Chinese sturgeon has become a critically endangered species. [0003] However, the research and protection of the genetic diversity of Chinese sturgeon requires a large number of samples of Chinese sturgeon for biological analysis, and the basis of biological analysis is often to extract DNA and RNA for research. The traditional method of extracting Chinese sturgeon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 胡亚成杜合军刘雪清肖衎刘娟娟陈冬娟王彬忠赵珣
Owner CHINESE STURGEON RES INST CHINA THREE GOR
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