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Excrement microbial sample DNA preserving fluid

A technology of fecal microorganisms and preservation solution, applied in the field of biological sample preservation solution, which can solve the problems of complex operation, poor applicability, and inapplicability of DNA preservation

Inactive Publication Date: 2017-05-31
SUZHOU PRECISION BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For the preservation of fecal microorganisms in the prior art, low-temperature freezing methods are often used. For example, the invention patent of Chinese Patent Publication No. CN102864138A discloses a method for extracting DNA from human feces. at -20°C to -80°C, however, the low-temperature freezing method requires the use of large-scale equipment, which has poor applicability and complicated operation, and is not suitable for extensive clinical testing; common DNA preservation solutions usually contain EDTA, although it can Used at room temperature, however, this DNA preservation solution is not suitable for DNA preservation of fecal microbial samples, and has limitations in use

Method used

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  • Excrement microbial sample DNA preserving fluid
  • Excrement microbial sample DNA preserving fluid
  • Excrement microbial sample DNA preserving fluid

Examples

Experimental program
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Effect test

Embodiment 1

[0026] 1. Sample source and group storage method

[0027] Fecal samples were collected from 3 volunteers. 9 samples were collected from each volunteer, and 9 different treatments were performed respectively:

[0028] Freeze for one week (label 0): Collect at least 0.2g of feces samples, quick-freeze in liquid nitrogen, and store in a sealed container at -80°C. After one week of storage, perform DNA extraction, 16S rRNA gene V3 / V4 variable region amplification, library construction, and sequencing Waiting for treatment;

[0029] Preservation solution for one week (label 1-7): collect at least 0.2g of feces samples, use 7 different formulas of the present invention (the ingredients are shown in Table 1) to seal at room temperature (25°C), store in the dark for one week, and then perform DNA extraction , 16S rRNA gene V3 / V4 variable region amplification, library construction, sequencing and other processing;

[0030] Store for one week without any treatment (label 8): Collect ...

Embodiment 2

[0059] 1. Sample source and group storage method

[0060] Fecal samples were collected from 3 volunteers. Four samples were collected from each volunteer, and they were treated in four different ways:

[0061] Freeze for one week: Collect at least 0.2g of feces samples, freeze them in liquid nitrogen, and store them sealed at -80°C. After one week of storage, perform DNA extraction, 16S rRNA gene V3 / V4 variable region amplification, library construction, sequencing, etc.;

[0062] Preservation of formula A for one week: Collect at least 0.2g of feces samples, use formula A (the ingredients are shown in Table 2) to seal at room temperature (25°C), store in the dark for one week, and then perform DNA extraction, 16S rRNA gene V3 / V4 variable region Amplification, library construction, sequencing and other processing;

[0063] Formula B is stored for one week: collect at least 0.2g of feces samples, use Formula B (the ingredients are shown in Table 3) to seal at room temperature...

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Abstract

The invention relates to biological sample preserving fluid, in particular to excrement microbial sample DNA preserving fluid. Ethylene diamine tetraacetic acid disodium (EDTA-2Na) of 0.15 to 0.35 mol / L, sodium chloride (NaCl) and dimethyl sulfoxide (DMSO) with the mass percent being 12 to 28 percent are dissolved in the excrement microbial sample DNA preserving fluid provided by the invention, and the pH value is adjusted to be 6.0 to 9.0. The excrement microbial sample DNA preserving fluid provided by the invention is suitable for preserving excrement microbial samples at normal temperature, is simple to operate and high in applicability, and can be applied to large-scale sample detection.

Description

technical field [0001] The invention relates to a biological sample preservation solution, in particular to a DNA preservation solution of a stool microbial sample. Background technique [0002] There are 10 trillion to 100 trillion bacteria in the human intestine, belonging to hundreds to thousands of different species, the number of which is 10 times the number of human cells, and the number of genes is 150-200 times that of human genes. These symbiotic microorganisms in the human intestinal tract are a large undiscovered "organ" of the human body. Some scholars believe that the human body itself is a symbiotic organism composed of human cells and bacteria. They live symbiotically with the human body, feed on the food ingested by the human body and intestinal secretions, synthesize essential amino acids, vitamins, short-chain fatty acids and other substances into the human body, and promote the health of the host; on the contrary, some of these pathogenic bacteria also sec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6806C12Q2527/125C12Q2527/119
Inventor 朱永亮
Owner SUZHOU PRECISION BIOTECH CO LTD
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