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Method for extracting microbial genome DNA (Deoxyribose Nucleic Acid) of frozen archenteron

An extraction method and genome technology, which is applied in the field of extraction of microbial genomic DNA in frozen original intestine, can solve the problems that the extraction of microbial genomic DNA cannot be fully applied, the content of animal intestinal tissue is large, and the number of bacteria is reduced, so as to save manpower, The test period is short and the effect of removing the interference of pollutants

Inactive Publication Date: 2014-08-13
NANTONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF THE PEOPLES REPUBLIC OF CHINA +1
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Problems solved by technology

The difference is that the frozen original intestine not only contains very little feces but more animal intestinal tissue, and the number of bacteria in the original intestine will decrease significantly during the process of removing feces and cleaning. Therefore, the above method cannot be fully applied to freezing Extraction of Microorganism Genomic DNA in Gastric

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  • Method for extracting microbial genome DNA (Deoxyribose Nucleic Acid) of frozen archenteron
  • Method for extracting microbial genome DNA (Deoxyribose Nucleic Acid) of frozen archenteron

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Embodiment Construction

[0030] Tween80 (Tween 80), Tris (tricarboxymethylaminomethane), EDTA (ethylenediaminetetraacetic acid) and Tris-saturated phenol (Tris-phenol, pH 7.8-8.0) are domestic reagents, NaC, absolute ethanol, Chloroform and isoamyl alcohol are all domestic analytical pure chemical reagents, the PCR kit (16S rDNA Bacterial Identification PCR Kit) is a product of Bao Biological Engineering (Dalian) Company (article number: D310), and the metal filter is from Daoxu Zhang, Shangyu City, Zhejiang Province. Products of Xing Yarn Sieve Factory.

[0031] (1) Take out the sealed and frozen sheep gastritis sample from the refrigerator, thaw it at room temperature, and use high-pressure sterilized scissors and tweezers in the ultra-clean workbench to cut the gastroenterum longitudinally and then fully cut it into pieces. Stir repeatedly to mix.

[0032] (2) Weigh 10 g of the mixed sample into a sterile 250 mL Erlenmeyer flask, then add 10 mL of sterilized normal saline (containing 0.1% Tween80)...

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Abstract

The invention discloses a method for extracting microbial genome DNA (Deoxyribose Nucleic Acid) of frozen archenteron. According to the method, a relatively large amount of samples (10g) are subjected to pretreatment by using a method with the combination of artificial crushing, eluting based on oscillation, filtration based on a metal net and gradient centrifugation before cell lysis. By adopting the method, cells are enriched, the problem that the content of microorganisms in the samples is relatively low is solved, the interference of pollutants is effectively prevented, and the DNA extraction efficiency and the DNA extraction purity are improved. The method is relatively good in university, the OD260 / OD230 and OD / 260 / OD280 of the microbial genome DNA of the microbial genome are approximate to standard values, and thus the DNA can be directly subjected to gene amplification and microbial diversity analysis.

Description

technical field [0001] The invention relates to a method for extracting microbial genome DNA in frozen original intestine. Background technique [0002] The original intestine (intestine) is the small intestine of unscraped healthy livestock, which is one of the important raw materials for the production of natural casings and used to make sausages. In order to prevent spoilage, original intestine must be preserved and transported under frozen conditions, so it is also called frozen original intestine. At present, the frozen original intestines used in the production and processing of natural casings at home and abroad are mainly small intestines of pigs and sheep. Frozen raw sausages come from animal intestines. After removing feces, cleaning and packaging in slaughterhouses, they are transported to casing companies for production and processing. Therefore, it is inevitable that frozen raw sausages will contaminate the microorganisms in and outside the intestines. The tota...

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Application Information

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IPC IPC(8): C12N15/10
Inventor 仇保丰宋鸿雁陈峰邵义祥刘文斌顾炳泉高逢结李建
Owner NANTONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF THE PEOPLES REPUBLIC OF CHINA