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Kit for easy and odorless extraction of fecal nucleic acid and extraction method

A kit and feces technology, applied in DNA preparation, recombinant DNA technology and other directions, can solve the problems of inconvenient transportation and short storage time, and achieve the effects of eliminating odor, using safety, and solving transportation and storage difficulties.

Active Publication Date: 2018-12-14
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a simple and odorless kit for extracting stool nucleic acid, which can effectively preserve the DNA in the stool sample for a long time at room temperature, and effectively solve the problem of short storage time of stool sample DNA and inconvenient transportation. Deficiency, which is conducive to the detection of nucleic acid in stool samples

Method used

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  • Kit for easy and odorless extraction of fecal nucleic acid and extraction method
  • Kit for easy and odorless extraction of fecal nucleic acid and extraction method
  • Kit for easy and odorless extraction of fecal nucleic acid and extraction method

Examples

Experimental program
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Effect test

preparation example Construction

[0019] The preparation method of natural green tea extract essence, comprises the steps:

[0020] Weigh the green tea leaves and purified water with a weight ratio of (1.5~2.3):(3.5~5.5), then mix the green tea leaves and purified water, and soak for 36~48h under the condition that the vacuum degree is -0.050~-0.060MPa, Reheat and decoct for 2 to 3 hours, and filter to obtain the filtrate and filter residue. Decolorize the filtrate, and concentrate the filtrate according to the volume ratio of (90-110):1 (for example, 90-110 L before concentration and 1 L after concentration) to obtain green tea extract.

[0021] The preparation method of honeysuckle extract comprises the following steps:

[0022] The honeysuckle medicinal material was pulverized and extracted with a 50% (v / v) ethanol aqueous solution to obtain an extract. The extract that obtains is concentrated under reduced pressure to obtain medicinal extract, add water to dissolve, filter, and solution is concentrated t...

no. 1 example

[0039] The simple and odorless kit for extracting stool nucleic acid provided in this example is as follows.

[0040] The kit includes stool stabilizing solution, lysate, first washing solution, second washing solution, eluent and chromatographic column.

[0041] Specifically, the stool stabilization liquid contains 8mM Tris, 2M guanidine thiocyanate, 6mM ethylenediaminetetraacetic acid, 0.05mM sodium chloride, 0.5% chlorobutanol, 8% (m / m) green tea extract, 6% ( m / m) honeysuckle extract and 4% (m / m) enzyme.

[0042] The lysate contains 15mmol / L Tris-HCl (pH 3.3), 10mmol / L guanidine isothiocyanate, 5mmol / L sodium hydroxide, 85% (v / v) isopropanol, 4% (v / v) Triton X-100, 2% (m / m) catechins and 0.5 μg / mL CarrierRNA.

[0043] The first washing solution contained 65 mmol / L Tris-HCl, 50% (v / v) ethanol and 0.5% (m / m) DTT.

[0044] The second washing solution contains 30mmol / L sodium acetate solution and 75% ethanol.

[0045] The eluate contains 30mmol / L Tris-HCl (pH 8), 3mmol / L E...

no. 2 example

[0048] This example provides a simple and odorless method for extracting stool nucleic acid.

[0049] (1) Human genomic DNA from stool samples

[0050] Stool sample processing steps

[0051] Take 0.2-0.4g of stool sample into a 2mL centrifuge tube. If the sample is in a liquid state, draw 0.2mL of the liquid sample into a 2mL centrifuge tube, add 0.65mL of the stool stabilization solution provided in the first embodiment to the sample, and vortex at the highest 1 minute to break up the sample. The samples can be stored in the stable solution for 15 days at room temperature, which is convenient for actual transportation and does not affect the subsequent detection of the samples.

[0052] protein cleavage step

[0053] Centrifuge at 13,000ⅹg for 10 minutes at room temperature. Then, transfer 500 μL of the supernatant to a 2 mL centrifuge tube, and add 20 μL of Proteinase K and the lysate provided in the first example to the supernatant. Inverted and mixed to obtain a mixtu...

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Abstract

The invention discloses a kit for easy and odorless extraction of fecal nucleic acid and an extraction method and relates to the field of molecular biotechnology. The kit includes a fecal stabilizingsolution containing Tris, guanidinium thiocyanate, ethylenediaminetetraacetic acid, sodium chloride, chlorobutanol, green tea extract, honeysuckle extract and an enzyme. The fecal stabilizing solutioncan store the DNA in the extracted fecal sample at a normal temperature of less than or equal to 40 DEG C for about 15 days, solve the problem of difficulty in transportation and storage and facilitate self-extraction and saving of the sample at home and is convenient and quick. The fecal stabilizing solution has the advantages of good deodorizing effect and use safety and can effectively removefecal odor gas such as ammonia gas and hydrogen sulfide thereby eliminating the odor and inhibiting the growth of bacteria.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a simple and odorless kit for extracting nucleic acid from feces and a method thereof. Background technique [0002] Stool, as a common clinical sample, contains a large amount of nucleic acid information. Due to the rhythmic nature of defecation, compared with other non-destructive sampling materials, the sample is easy to collect and obtain. Therefore, stool samples can be used for the detection of related tumors, the most A common use is in the diagnosis of colorectal cancer. [0003] However, after the excrement of feces, its environment has also changed, mainly from an anaerobic environment to an aerobic environment. At the same time, some bacteria in it still maintain some physiological activity and continue to reproduce and metabolize. However, due to changes in the environment, the growth and death rates of different bacteria are different from those before def...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 许嘉森吴诗扬刘志明赖炳林
Owner SUREXAM BIO TECH
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