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Simple method for efficiently extracting DNA (deoxyribonucleic acid) from soil samples

A soil sample, high-efficiency technology, applied in the field of environmental science and bioengineering, can solve the problems of high price, low extraction efficiency, and small amount of DNA extraction, and achieve the effect of strong practicability and high reliability

Active Publication Date: 2014-08-20
NANKAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the problems of the existing microbial DNA extraction methods, such as small amount of extracted DNA, low purity, low extraction efficiency, expensive reagents, etc., and propose a simple and efficient method for extracting DNA

Method used

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  • Simple method for efficiently extracting DNA (deoxyribonucleic acid) from soil samples
  • Simple method for efficiently extracting DNA (deoxyribonucleic acid) from soil samples
  • Simple method for efficiently extracting DNA (deoxyribonucleic acid) from soil samples

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Experimental program
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Embodiment 1

[0051] Embodiment 1: the extraction of DNA in the soil sample;

[0052] (1) Cultivation of internal standard bacteria Xcc8004;

[0053] Inoculate Xcc8004 bacteria into NYG liquid medium, place in a constant temperature shaking incubator at 28°C for shaking culture for 20 hours; use the spread plate method or flow cytometry to determine the concentration of the bacteria solution;

[0054] Wherein, the formula of NYG liquid culture medium is: the peptone that mass fraction is 0.5%, the yeast extract that mass fraction is 0.3%, the glycerin that mass fraction is 2%, and the solid medium that agar powder mass fraction is 1%, adjust The pH of the culture medium is 7.0; the preparation concentration of rifampicin antibiotic mother solution is 10 mg / ml, the solvent is methanol, and the action concentration is 50 μg / ml;

[0055] The microscope morphology observation of internal standard bacteria Xcc8004 is attached figure 1 shown;

[0056] (2) Lysis of microbial cells in soil sampl...

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Abstract

The invention provides a simple method for efficiently extracting DNA (deoxyribonucleic acid) from soil samples. According to the simple method, the DNA in environmental samples is fully released and exposed by using combination of methods such as glass bead grinding, liquid nitrogen repetitive freeze-thawing, SDS (sodium-dodecyl sulphate), lysozyme and a protease-K method for the different soil samples, the DNA purification is carried out through multiple phenol imitation extraction processes, and the determination of the DNA extraction efficiency is carried out by adding an internal standard bacterial strain Xcc8004. The simple method has simple requirements on sample pretreatment; the quantity of the required environmental samples is small; the obtained DNA has relatively high concentration and purity; the extraction efficiency is relatively high; the analysis and detection of antibiotic resistance gene pollution in the complex environmental samples can be met; and the method is simple and reliable and has practical application value.

Description

technical field [0001] The invention relates to a simple method for extracting microbial DNA from environmental samples, especially soil samples. The method processes the environmental soil samples by means of biotechnology, and belongs to the technical fields of environmental science and bioengineering. Background technique [0002] The widespread existence and spread of antibiotic resistance genes in the natural environment has caused huge environmental and health risks. It is necessary to accurately detect the number of resistance genes in the natural environment. Soil medium is a place where many microorganisms inhabit in nature. At present, many studies have been carried out on the diversity of soil microorganisms and the detection of the number of resistance genes at the DNA molecular level. The DNA extraction process of soil samples mainly includes cell wall lysis, protein and polysaccharide removal, and DNA precipitation recovery. However, due to the complex soil m...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12R1/64
Inventor 王记鲁毛大庆罗义
Owner NANKAI UNIV
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