Method for extracting microorganism total DNA (Deoxyribonucleic Acid) in pu'er tea piling fermentation process

Abstract

The invention discloses a method for extracting microorganism total DNA (Deoxyribonucleic Acid) in a pu'er tea piling fermentation process at high quality. The method comprises the following steps of: separating microorganisms on the surface of pu'er tea by using ultrasonic wave and vortex oscillation; washing the microorganisms by using washing buffer solution; performing rough extraction on the microorganism total DNA by using DNA extract; and purifying the roughly extracted DNA. The method provided by the invention is low in cost and high in DNA extraction rate; the extracted DNA has high completeness and high purity and can meet the requirements of PCR (Polymerase Chain Reaction) without purification or dilution; the purified DNA can meet the requirements of molecular biology research; and meanwhile, the extracted DNA covers bacteria and fungi and the requirements of subsequent researches in microorganism diversity, functional genes, metagenomics and the like can be met.

Description

technical field [0001] The invention relates to a method for extracting the total DNA of microorganisms in the fermentation process of Pu'er tea, which belongs to the field of microorganisms. Background technique [0002] Pu-erh tea is a large leaf tea from Yunnan ( C. sinensis var. assamica ) and other large-leaf teas (Pu-erh green tea) as raw materials, loose tea and pressed tea produced by post-fermentation process. Stacking fermentation in the modern Pu'er tea production process is a key process for producing Pu'er tea. The types and quantities of microorganisms in the production process directly affect the formation of Pu'er tea flavor. Therefore, microorganisms play an important role in the stacking fermentation of Pu'er ripe tea. effect. However, the current detection and classification methods based on morphological and physiological and biochemical indicators are limited by culture conditions and simulated culture techniques, so there is still a big difference bet...

AI Technical Summary

Problems solved by technology

[0003] At present, the main problem in the extraction of the total microbial DNA of Pu’er tea is that the microorganisms use Pu’er tea as a substrate to perform complex secondary metabolism, and a large amount of tea polyphenols, tea polysaccharides, quinones, etc. remain on the surface and even in the body of the microorganisms. Makes it very difficult to extract high-quality microbial total DNA

According to the current method, the microbial total DNA of Puer tea is extracted. It is difficult to avoid the combination of tea polyphenols and quinones with DNA during the extraction process. The extracted DNA has a high content of impurities and cannot be directly used for subsequent molecular biology research such as PCR amplification. The current method still has insufficient cell lysis rate in the extraction process, and the combination of DNA and tea polyphenols, quinones, etc. leads to excessive loss in the extraction process, etc., resulting in a low extraction rate

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