Method for extraction and purification of microbial total DNA of compost and buffer solution thereof

A purification method and microbial technology, applied in the direction of DNA preparation, recombinant DNA technology, sugar derivatives, etc., can solve the problems affecting the quantitative molecular biology operation process of nucleic acid, and achieve the goals of improving lysis efficiency, good integrity, and avoiding mechanical damage Effect

Inactive Publication Date: 2009-10-07
HUNAN UNIV
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Problems solved by technology

Even in ethanol or isopropanol solutions where DNA is purified and concentrated, humic acid still exhibits a similar precipitation behavior to DNA, so some physicochemical properties remain during further purification of crude nucleic acid extra

Method used

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  • Method for extraction and purification of microbial total DNA of compost and buffer solution thereof
  • Method for extraction and purification of microbial total DNA of compost and buffer solution thereof
  • Method for extraction and purification of microbial total DNA of compost and buffer solution thereof

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Embodiment

[0038] Sampling of the compost was performed first. The compost was taken from a 20L experimental-scale composting device. The compost materials mainly included 1.074kg of straw, 0.9kg of vegetable leaves, 0.216kg of leaves, 0.24kg of bran and 0.721kg of soil. The sampling point was 3cm below the compost surface, and a total of three Samples (respectively numbered 1, 2, 3), each sample 1g, sampling time is 18 days after composting, temperature is 38 ℃, pH value is 8.32, moisture content is 58%, organic matter content is 20.3%. Among them, the pH value is measured by the glass electrode method, the moisture content is measured by repeated baking at 105°C until constant weight is measured, and the organic matter content is measured by the potassium dichromate method.

[0039] Adopt the method of the present invention to carry out the extraction and the purification of composting microbial total DNA to above-mentioned composting sample, specifically comprise the following steps: ...

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Abstract

The invention discloses a method for the extraction and the purification of microbial total DNA of a compost and a buffer solution thereof, the method comprises the following steps of: adding a dehumification solution in an amount of 8-15ml/g of the compost into a sample of the compost in which the DNA is to be extracted, for dehumification; then adding a DNA extraction buffer solution in an amount of 1.0-1.5ml/g of the compost for the crude extraction of the microbial total DNA; and finally, purifying the crudely-extracted DNA solution with a kit so as to obtain the microbial total DNA of the compost. The inventive method can reduce the influence of humic acids and effectively improve the yield and the purity of the DNA extracted from the compost, thus better facilitating compost microbial molecular ecology and researches of other functional genes.

Description

technical field [0001] The invention relates to a method for extracting and purifying DNA, in particular to a method for extracting and purifying total DNA of microorganisms and a used buffer. Background technique [0002] Composting refers to the process of artificially promoting the transformation of biodegradable organic matter in solid waste into stable humus by utilizing the mutual synergy of microorganisms such as bacteria, fungi, and actinomycetes widely distributed in nature. Therefore, fully understanding the microbiological principles It can provide a basis for improving the treatment process and improving the treatment efficiency. The traditional method of microbial research on compost is mainly microbial culture and pure species isolation technology, because most of the microorganisms (>95%) in the environment cannot be studied by culture, and there are many kinds of microorganisms in compost, which are often in dynamic changes state, culture studies cannot r...

Claims

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Application Information

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IPC IPC(8): C12N15/10C07H21/04
Inventor 曾光明范长征杨春平汤琳李贞李凤肖勇
Owner HUNAN UNIV
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