Microbe sample preservation solution for gene detection, preparation method and kit thereof, and application of kit

A gene detection and microbial technology, applied in the field of bioengineering, can solve the problems of reduced inactivation effect, cell rupture, easy to be degraded and damaged, etc., and achieve the effect of comprehensive microbial inactivation ability, maintenance of cell structure, and thorough inactivation effect

Active Publication Date: 2018-09-18
知几未来(成都)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, the oral / fecal bacterial preservation solutions on the market are mainly composed of ethanol / methanol / SDS, and are prepared into weakly alkaline isotonic solutions, which can kill the bacteria in the sample, but will cause damage to the cell structure, resulting in Cell contents are released and cannot be used for immunological or other methods of detection
At the same time, methanol\guanidine isothiocyanate in the preservation solution is volatile or highly toxic, and it is easy to cause volatilization during use and cause harm to operators
[0003] The existing preservation solution is inactivated with high-purity ethanol; some bacteria will have a stress response to high-concentration ethanol, resulting in a reduced inactivation effect; it contains protective nucleic acid components, but a large amount of surfactant in the preservation solution can also cause cell rupture , causing nucleic acid etc. to be in an exposed environment and easily degraded and destroyed; some inactivators are highly toxic, and operational errors during use will cause poisoning to users

Method used

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  • Microbe sample preservation solution for gene detection, preparation method and kit thereof, and application of kit
  • Microbe sample preservation solution for gene detection, preparation method and kit thereof, and application of kit
  • Microbe sample preservation solution for gene detection, preparation method and kit thereof, and application of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0036] The present embodiment provides a microbial sample preservation solution for genetic testing. The microbial sample preservation solution includes DMSO 17mL, glycerol 0.7mL, edetate disodium 8.2g, NaCl 2.5g, guanidine hydrochloride, guanidine hydrochloride 2.7g and Tris HCl. .8g.

[0037] This embodiment also provides a method for preparing a microbial sample preservation solution for genetic testing, comprising the following steps:

[0038] 1.1 Measure 20mL of DMSO and 0.7mL of glycerin, weigh 8.2g of disodium edetate, 2.5g of NaCl, guanidine hydrochloride, 2.7g of guanidine hydrochloride and 0.8g of Tris·HCl, and mix them in a beaker;

[0039] 1.2 Add 45mL of absolute ethanol to the beaker to dissolve;

[0040] 1.3 Dilute to 100mL with double distilled deionized water;

[0041] 1.4 After mixing, adjust the pH value to 7.3 with hydrochloric acid.

[0042] The microbial sample preservation solution for genetic testing is prepared, so the microbial sample preservation ...

Embodiment 2

[0046] This example provides a microbial sample preservation solution for genetic testing. The microbial sample preservation solution includes 20 mL of DMSO, 0.9 mL of glycerol, 3.2 g of guanidine hydrochloride, 9.1 g of disodium edetate, 2.7 g of NaCl and 1.0 g of Tris·HCl.

[0047] This embodiment also provides a method for preparing a microbial sample preservation solution for genetic testing, comprising the following steps:

[0048] 1.1 Measure 20mL of DMSO and 0.9mL of glycerin, weigh 9.1g of disodium edetate, 2.7g of NaCl, guanidine hydrochloride, 3.2g of guanidine hydrochloride and 1.0g of Tris HCl, and mix them in a beaker;

[0049]1.2 Add 50mL of absolute ethanol to the beaker to dissolve;

[0050] 1.3 Dilute to 100mL with double distilled deionized water;

[0051] 1.4 After mixing, adjust the pH value to 7.4 with hydrochloric acid.

[0052] The microbial sample preservation solution for genetic testing is prepared, so the microbial sample preservation solution can ...

Embodiment 3

[0056] This embodiment provides a microbial sample preservation solution for genetic testing. The microbial sample preservation solution includes 24 mL of DMSO, 1.3 mL of glycerol, 12.5 g of disodium edetate, 3.7 g of NaCl, 4.1 g of guanidine hydrochloride and 1.3 g of Tris·HCl.

[0057] This embodiment also provides a method for preparing a microbial sample preservation solution for genetic testing, comprising the following steps:

[0058] 1.1 Measure 24mL of DMSO and 1.3mL of glycerin, weigh 12.5g of disodium edetate, 3.7g of NaCl, guanidine hydrochloride, 4.1g of guanidine hydrochloride and 1.3g of Tris HCl, and mix them in a beaker;

[0059] 1.2 Add 52mL of absolute ethanol to the beaker to dissolve;

[0060] 1.3 Dilute to 100mL with double distilled deionized water;

[0061] 1.4 After mixing, adjust the pH value to 7.5 with hydrochloric acid.

[0062] The microbial sample preservation solution for genetic testing is prepared, so the microbial sample preservation solutio...

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Abstract

The invention provides a microbe sample preservation solution for gene detection, a preparation method and a kit thereof, and an application of the kit, and belongs to the technical field of bioengineering. The microbe sample preservation solution for gene detection has effective and comprehensive microbe inactivation capability; and low-concentration ethanol is used to thoroughly inactivate bacteria in the storage process of a sample, and avoids inactivation failure caused by the stress reaction of bacteria due to high-concentration ethanol. The microbe sample preservation solution has good permeability on the premise of not destroying the cell structure, and effectively destroys the protein activity of bacteria and viruses and the activities of various nucleases to make the bacteria, viruses and nucleases not have of infection or proliferation ability, so the thorough inactivation effect is achieved. A glycerol-containing cytoprotective agent and a physiological concentration buffersolution are used to keep the cell structure of microbes and prevent the cell structure from be ruptured in order to protect the integrity of nucleic acids in cells; and the microbe sample preservation solution contains no high-toxicity components or volatile components, so no volatile poisoning occurs during use.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a microbial sample preservation solution for gene detection, a preparation method, a kit and an application thereof. Background technique [0002] At present, the oral / fecal bacterial preservation solutions on the market are mainly composed of ethanol / methanol / SDS, and are prepared into weakly alkaline isotonic solutions, which can kill the bacteria in the sample, but will cause damage to the cell structure, resulting in Cell contents are released and cannot be detected by immunological or other methods. At the same time, methanol\guanidine isothiocyanate in the preservation solution is volatile or highly toxic, and it is easy to cause volatilization during use and cause harm to operators. [0003] The existing preservation solution is inactivated with high-purity ethanol; some bacteria will have a stress response to high-concentration ethanol, resulting in a reduced inac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 陈裕殷高芸夏广亮廖洁莹索荔黄卫娟潘伟颜红眭麟
Owner 知几未来(成都)生物科技有限公司
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