Kit and extraction method for rapidly extracting RNA of animal fecal microorganism

An animal feces and kit technology, applied in the biological field, can solve problems such as degradation and long extraction time, and achieve the effect of simplifying experimental steps and saving extraction time.

Inactive Publication Date: 2015-03-25
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few reports on the extraction of microbial RNA from animal feces, because the feces contain more complex substances, and the extraction time of traditional methods is too long, which often causes the degradation of the extracted fecal microbial RNA.

Method used

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  • Kit and extraction method for rapidly extracting RNA of animal fecal microorganism

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] A kit for rapidly extracting microbial RNA from animal feces, the kit consists of an adsorption column and the following working solution:

[0050] Working solution A: composed of potassium dihydrogen phosphate at a final concentration of 0.25 and sodium hydroxide at a final concentration of 0.175, pH=7.2;

[0051] Working solution B: composed of tris hydrochloride (Tris-HCl) at a final concentration of 10 mmol / L and ethylenediaminetetraacetic acid (EDTA) at a final concentration of 1 mmol / L, pH=8.0;

[0052] Working solution C: the mass concentration is 1% lysozyme;

[0053] Working solution D: 20mg / mL proteinase K;

[0054] Working solution E: sodium dodecyl sulfate (SDS) with a mass concentration of 10%;

[0055] Working solution F: a mixed solution with a volume ratio of phenol: chloroform: isoamyl alcohol = 25:24:1;

[0056] Working solution G: a mixed solution with a volume ratio of chloroform: isoamyl alcohol = 24:1;

[0057] Working solution H: volume concen...

Embodiment 2

[0062] A kit for rapidly extracting microbial RNA from animal feces, the kit consists of an adsorption column and the following working solution:

[0063] Working solution A: composed of potassium dihydrogen phosphate with a final concentration of 0.37mol / L and sodium hydroxide with a final concentration of 0.2mol / L, pH=8;

[0064] Working solution B: composed of tris hydrochloride (Tris-HCl) at a final concentration of 20mmol / L and ethylenediaminetetraacetic acid (EDTA) at a final concentration of 10mmol / L, pH=5;

[0065] Working solution C: the mass concentration is 2% lysozyme;

[0066] Working solution D: 10mg / mL proteinase K;

[0067] Working solution E: sodium dodecyl sulfate (SDS) with a mass concentration of 20%;

[0068] Working solution F: a mixed solution with a volume ratio of phenol: chloroform: isoamyl alcohol = 25:24:1;

[0069] Working solution G: a mixed solution with a volume ratio of chloroform: isoamyl alcohol = 24:1;

[0070] Working solution H: volume...

Embodiment 3

[0075] A kit for rapidly extracting microbial RNA from animal feces, the kit consists of an adsorption column and the following working solution:

[0076] Working solution A: composed of potassium dihydrogen phosphate with a final concentration of 0.30mol / L and sodium hydroxide with a final concentration of 0.18mol / L, pH=7;

[0077] Working solution B: composed of tris hydrochloride (Tris-HCl) at a final concentration of 15mmol / L and ethylenediaminetetraacetic acid (EDTA) at a final concentration of 5mmol / L, pH=6.5;

[0078] Working solution C: the mass concentration is 1.5% lysozyme;

[0079] Working solution D: 15mg / mL proteinase K;

[0080] Working solution E: sodium dodecyl sulfate (SDS) with a mass concentration of 15%;

[0081] Working solution F: a mixed solution with a volume ratio of phenol: chloroform: isoamyl alcohol = 25:24:1;

[0082] Working solution G: a mixed solution with a volume ratio of chloroform: isoamyl alcohol = 24:1;

[0083] Working solution H: vo...

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Abstract

The invention discloses a kit and an extraction method for rapidly extracting RNA of animal fecal microorganism. The kit mainly adopts a silicone mold adsorption column to absorb the RNA of the fecal microorganism, and then adopts a peculiar protein removing solution and a rinsing solution to wash away impurities such as proteins and the like. Compared with the conventional method for extracting the RNA of the animal fecal microorganism, the extracting method by using the kit saves plenty of time for extraction and meanwhile simplifies experimental procedure, and can extract the RNA of the animal fecal microorganism more effectively, rapidly and economically.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit and an extraction method for rapidly extracting animal feces microbial RNA. Background technique [0002] Among the traditional research methods of intestinal microbes, the microscope method is very valuable for counting the flora. This method is suitable for clinical application due to the simple equipment required, easy operation, and short time-consuming. However, this method cannot fully reflect the state of the intestinal flora, let alone accurately quantify the types and quantities of microorganisms, and the operation process is greatly affected by subjective factors. The most important point is that pure cultures cannot be obtained and cannot be analyzed. Further research has limitations. [0003] With the rapid development of molecular biology, more and more technologies have been applied to the study of gut microbes, and the two most important molecular su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 王清水余彦
Owner FUJIAN NORMAL UNIV
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