Total DNA extraction method and kit for simultaneous removal of humic acid and bacterial protein

Abstract

The invention provides a total DNA (deoxyribonucleic acid) extraction method and a kit for synchronously removing humic acid and mycoprotein and relates to the field of research of metagenomics in soil microbiology and general microbiology. The method comprises the following steps: (1) suspending samples by utilizing a humic acid combination solution, so as to obtain sediments; (2) adding a buffer solution and proteinase K, and carrying out centrifuging to obtain supernate; (3) adding a coagulant to carry out coagulating sedimentation on the residual humic acid and mycoprotein, and carrying out centrifuging to obtain the supernate; (4) adding isopropanol to precipitate DNA; (5) adding 70% ethanol to wash the sediments, and blowing off the residual ethanol; (6) adding a DNA dissolving solution (TE) to dissolve the sediments, so as to obtain a sample, namely the total DNA. The kit comprises the following components: the humic acid combination solution, a PBS butter liquid, an extraction butter liquid, the proteinase K, a 20% SDS (sodium dodecyl sulfonate) water solution, the coagulant, a DNA washing agent, the 70% ethanol and the DNA dissolving solution. Due to the adoption of the method and the kit, the one-time extraction quantity of the total DNA is enough to carry out PCR (polymerase chain reaction) amplification for dozens of times.

Description

Technical field [0001] The invention relates to the research field of metagenomics in soil microorganisms and general microbiology, and specifically is a simple and rapid method for extracting total DNA of soil microorganisms (including general microorganisms). Background technique [0002] In order to reveal the differences and characteristics of the structure, species and distribution of microbial flora in different ecosystems, 16SrRNA gene sequence analysis methods have been widely used. This requires that a qualified sample of total microbial DNA can be obtained first. Soil is a major part of the ecosystem, and soil microbial flora analysis is particularly important. Although the total DNA of microorganisms in the general environment is relatively easy to extract, the extraction of total DNA of soil microorganisms is affected by various soil components, especially The effect of humic acid is more difficult. Fortunately, there are many corresponding treatment measures. There ...

AI Technical Summary

Problems solved by technology

As a main part of the ecosystem, soil microbial flora analysis is particularly important. Although the total DNA of microorganisms in the general environment is relatively easy to extract, the extraction of total DNA of soil microorganisms is affected by various soil components, especially The impact of humic acid is more difficult to carry out. Fortunately, there are many corresponding treatment measures now, and there are various soil microbial total DNA extraction methods or kits, but they always have one or another deficiency, such as convenience and simplicity. lack of sex, economy, etc.

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