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

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...

AI Technical Summary

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 extracts Humic acid components that are similar to nucleic acids coexist with them, thus affecting the quantification of nucleic acids and subsequent molecular biology operations such as PCR amplification, hybridization, enzyme digestion, and cloning

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