Method for extracting microbial dna from lignocellulose anaerobic fermentation material and its complete set of reagents

Abstract

The invention discloses a method for extracting DNA of microorganisms on anaerobic fermentation materials of lignocelluloses and a complete set of reagent. The method comprises the following steps of (S1) washing microorganisms on materials by using an eluant to obtain a microorganism eluate, and removing impurities in the microorganism eluate to obtain microorganisms, wherein the eluant is composed of a solvent and a solute, the solvent is water, and the solute comprises the components with the concentrations as follows: 9.0g / L sodium chloride, 0.12g / L potassium chloride, 0.24g / L calcium chloride, 0.2g / L sodium bicarbonate and 0.15g / L monopotassium phosphate; and (S2) extracting the DNA of the microorganisms to obtain the DNA of the microorganisms. Experiments prove that method for extracting the DNA of the microorganisms on the anaerobic fermentation materials of lignocelluloses and the complete set of reagent can be used for extracting the DNA of microorganisms on anaerobic fermentation materials of different lignocelluloses.

Description

technical field [0001] The invention relates to a method for extracting microbial DNA of lignocellulose anaerobic fermentation materials and a complete set of reagents in the field of biotechnology. Background technique [0002] The essence of anaerobic fermentation methane production process is that multiple microbial groups work together to continuously decompose complex lignocellulose until methane and carbon dioxide are produced. In recent years, with the increasing shortage of natural gas energy and smog pollution, the advantages of bio-natural gas (biogas with methane content greater than 95%), such as low cost, clean environmental protection, and sustainability, have gradually emerged, and the process of producing methane from lignocellulosic materials has begun To the direction of scale, industrialization and industrialization. Qualitative and quantitative understanding of the diversity and growth and reproduction rules of each functional microbial group in the anae...

AI Technical Summary

Problems solved by technology

Although the traditional SDS method, CTAB method and SDS-CTAB combination method extract high quality and purity of total DNA, it is difficult to effectively obtain microbial DNA adsorbed on the surface of lignocellulose, resulting in the loss of some key microbial group information, and The number of samples taken at one time is small

Functional kit methods such as soil, silt, and feces have the advantages of strong specificity, rapidity and convenience, and a large sample size for one extraction. However, for lignocellulosic anaerobic fermentation materials with high humic acid and miscellaneous protein content, The impurity removal ability of this method is limited, and the total DNA purity of the extracted microorganisms is relatively poor; in addition, the sample quality required for this method to extract the total DNA at one time is very little (0.3-0.5g), and the concentration of the total microbial DNA extracted is also low

At present, it is difficult for most methods to obtain total microbial DNA with high purity and concentration for lignocellulosic materials. There is an urgent need for a mature method for extracting microbial DNA from lignocellulosic anaerobic fermentation materials.

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