Method for extracting total DNA (Deoxyribonucleic Acid) of microorganisms from pile-fermentation Pu-erh tea

A compost fermentation and microbial technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of uncultivable microorganisms and achieve the effect of short experiment time

Active Publication Date: 2012-07-11
TIANJIN TASLY GROUP
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  • Claims
  • Application Information

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Problems solved by technology

The obtained microbial total DNA can comprehensively include the microbial species in the Pu'er tea fermentation process, which can overcome the defect that the traditional microbial isolation technology cannot obtain unculturable microorganisms

Method used

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  • Method for extracting total DNA (Deoxyribonucleic Acid) of microorganisms from pile-fermentation Pu-erh tea
  • Method for extracting total DNA (Deoxyribonucleic Acid) of microorganisms from pile-fermentation Pu-erh tea
  • Method for extracting total DNA (Deoxyribonucleic Acid) of microorganisms from pile-fermentation Pu-erh tea

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Sampling of pile-fermented Pu-erh tea, taking pile-fermented Pu-erh tea with different turning times and different stacking positions, analyzing the population structure of bacteria and determining the dominant flora are as follows

[0033] 1) Take 5g tea sample, use liquid nitrogen to freeze and grind 3 times, transfer to a 2ml centrifuge tube, add 200mg glass beads. Use the Omega soil DNA extraction kit to extract the treated tea samples, add 600ul SLX solution, mix evenly with high-speed shaking for 10 minutes, and bathe in 70°C water for 10 minutes. During this period, mix the samples several times, add 200ul SP2 solution, mix well, and ice bath for 5 minutes. Centrifuge at 13000g for 5min. Transfer the supernatant to a new centrifuge tube, add 0.7 times isopropanol, invert and mix well, centrifuge at 13000g for 10min, discard the supernatant, invert on absorbent paper to dry, add 200ul elution buffer to the centrifuge tube, mix well, Dissolve DNA in a water bath a...

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Abstract

The invention relates to a method for extracting total DNA (Deoxyribonucleic Acid) of microorganisms from pile-fermentation Pu-erh tea. The method comprises the following steps of: 1) taking Pu-erh tea samples in different fermentation processes; 2) pretreating the Pu-erh tea samples to be extracted; 3) extracting the total DNA of the microorganisms from the pretreated samples; 4) carrying out agarose gel electrophoresis detection on the extracted total DNA of the microorganisms; 5) carrying out PCR (Polymerase Chain Reaction) amplification under the guidance of special primers by using the extracted total DNA as a template; and 6) carrying out the agarose gel electrophoresis detection on the fragments subjected to the PCR amplification.

Description

Technical field: [0001] The invention relates to a method for extracting and detecting microbial total DNA in pile-fermented Pu'er tea, belonging to the field of microbial ecology. Background technique: [0002] Pu-erh cooked tea is a post-fermentation made from the sun-dried green tea of ​​Yunnan's unique big-leaf tea [Camellia sinensis (Linn) var. tea. Microorganisms play an important role in the fermentation process of Pu'er tea. Without microorganisms, the quality and flavor of Pu'er tea will not be guaranteed. The present invention studies the dominant microbial flora in Pu'er tea fermented by stacking. [0003] At present, the research on microorganisms in fermented Pu-erh tea mainly uses traditional methods such as separation, purification and identification of microorganisms. The traditional research method is to first sample from a specific sample, use various media and conditions that simulate the natural environment for isolation and culture, and obtain strains ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 李长文李巍梁慧珍张长霞李季刘冰闫希军
Owner TIANJIN TASLY GROUP
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