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Preservation reagent for microorganism specimens containing rich organic matters, kit containing preservation reagent and application of preservation reagent

A technology for preserving reagents and organic matter, which is applied in the field of preservation reagents for microbial samples, and can solve problems such as difficulty in implementation and harsh conditions for cryogenic freezing.

Inactive Publication Date: 2016-05-04
TINYGENE BIOTECH SHANGHAI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, its biggest drawback is that the conditions for cryopreservation are harsh and difficult to achieve.

Method used

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  • Preservation reagent for microorganism specimens containing rich organic matters, kit containing preservation reagent and application of preservation reagent
  • Preservation reagent for microorganism specimens containing rich organic matters, kit containing preservation reagent and application of preservation reagent
  • Preservation reagent for microorganism specimens containing rich organic matters, kit containing preservation reagent and application of preservation reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Preparation of preservation reagents for microbial samples rich in organic matter.

[0028] According to the formula, first prepare a buffer solution with a suitable concentration with deionized water, then add an appropriate amount of cell protection agent, chelating agent and ionic strength maintenance agent according to the volume ratio, put it on a magnetic stirrer, and mix well, so that the final The pH value of the pH buffer solution in the preservation reagent is 7.0-8.5, the concentration of the microbial cell protection agent is 5%-40%, the concentration of the chelating agent is 100-250mM, and the concentration of the ionic strength maintaining agent is not lower than 10mM. Store at room temperature.

Embodiment 2

[0029] Example 2: Collection of stool samples and treatment of different storage times.

[0030] Take the fresh feces of two volunteers (the middle part is excreted after taking, avoiding the mucous membrane on the surface of the feces, which can better reflect the intestinal microbial groups), and divide them into 12 tubes, of which 3 tubes are immediately frozen at -80 ℃ , as a parallel experiment and a positive control; add the preservation reagent described in the invention to the remaining 9 tubes, place them in a 37°C water bath, and store them at a constant temperature for 3 days, 7 days, and 14 days. Immediately after sampling, the samples were frozen and stored in a -80°C refrigerator for subsequent genome extraction.

Embodiment 3

[0031] Example 3: Experiment on preservation effect of bacterial total DNA in stool samples.

[0032] Using QIAampDNAStoolMiniKit for genome extraction, electrophoresis detection for quality control, the results are as follows Figure 1a and 1b As shown, the concentration is 10-20ng / uL.

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Abstract

The invention discloses a preservation reagent for microorganism specimens containing rich organic matters, a kit containing the preservation reagent and an application of the preservation reagent. The preservation reagent comprises a buffer solution, a microorganism cell protective agent, a chelating agent and an ion strength maintenance agent. The preservation reagent contains simple and easy-to-prepare ingredients, is simple and convenient to prepare and low in cost, has a good preservation effect on microorganisms in the specimens with rich organic matters, can guarantee the diversity and the corresponding proportion of the microorganisms in the specimens at the normal temperature or even at the higher temperature, maintains the microorganism unchanged in at least 14 days, is applicable to outdoor sampling and the conditions that immediate frozen storage is not convenient after sampling or frozen storage is unavailable, has lower requirements for the transportation conditions and facilitates storage and transportation of the specimens. With the use of the preservation reagent, storage of the specimens containing the rich organic matters is facilitated, no influence is produced on following extraction of total DNA, PCR amplification, high-throughput sequencing and the like of microorganisms, and the experiment efficiency can be further improved.

Description

technical field [0001] The present invention relates to the field of microbial molecular ecology, in particular to a preservation reagent for microbial samples rich in organic matter, a kit containing the preservation reagent and its application; the present invention also relates to the use of the preservation reagent for microbial samples rich in organic matter And detection kits and methods. Background technique [0002] Microorganisms are particularly abundant in organic matter-rich environments, both in quantity and variety. Since the environment is rich in organic nutrients, after the sample is collected, the microorganisms in the sample can still use the nutrients in it to continue to grow and reproduce. However, since the environment of the sample has been changed after sampling, at this time, if the microorganisms in the sample continue to grow and reproduce, the microflora structure of the microorganisms will change. Certain groups of microorganisms are more adap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 丁旭侯美玲原雅纬
Owner TINYGENE BIOTECH SHANGHAI CO LTD
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