RNA extraction solution, method for preparing RNA extraction solution, and RNA reagent extraction method

A solution and total amount technology, applied in the field of biomedicine, can solve the problems of increased labor, reduced RNA extraction, RNA loss, etc., and achieves the effects of avoiding impurities mixing, preventing acid hydrolysis, and simplifying operation steps.

Inactive Publication Date: 2019-01-18
SUZHOU NEW CELL & MOLECULAR BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the case of liquid treatment with DNase, in order to remove DNase after treatment, it is necessary to perform phenol / chloroform extraction or protein denaturation experiment again
In addition, it can be processed in combination with a silica membrane adsorption column, but the steps of washing the adsorption column must be repeated
These methods not only increase the amount of labor when removing impurity DNA, but also cause the loss of RNA, thereby reducing the amount of RNA extracted.

Method used

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Examples

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preparation example Construction

[0030] On the basis of the above, the present invention also discloses a method for preparing an RNA extraction solution, comprising the following steps: assuming that the total amount of the RNA extraction solution is 1000ml;

[0031] (1) Preparation of guanidine isothiocyanate solution: take 200ml ultrapure water as a solvent, slowly add 50g-250g of guanidine isothiocyanate, stir evenly on a magnetic stirrer to fully dissolve it;

[0032] (2) Measure the glycerin solution of 30-100ml, join in the above-mentioned guanidine isothiocyanate solution, carry out sufficient stirring;

[0033] (3) Add 0.1%-1% sodium lauryl sarcosinate to the above solution, and stir evenly;

[0034] (4) Add acetic acid solution and sodium acetate solution with a concentration of 0.05-0.2M relative to the total amount of the solution respectively, and the volume ratio is 3:1-5:1 to the above-mentioned solution, and fully stir evenly;

[0035] (5) Add 30-50% volume of hot saturated phenol solution to t...

Embodiment example 1

[0050] Preparation of high-purity RNA extraction solution:

[0051] The total amount of high-purity RNA extraction solution is 1000ml;

[0052] (1) Preparation of guanidine isothiocyanate solution: take 200ml ultrapure water as a solvent, slowly add 250g of guanidine isothiocyanate, stir evenly on a magnetic stirrer to fully dissolve it;

[0053] (2) Measure the glycerin solution of 50ml, join in the above-mentioned guanidine isothiocyanate solution, carry out sufficient stirring;

[0054] (3) Add 0.5% sodium lauryl sarcosinate relative to the total amount of the solution to the above solution, and stir evenly;

[0055] (4) Add acetic acid solution and sodium acetate solution with a concentration of 0.1M relative to the total amount of the solution, respectively, into the above solution at a volume ratio of 4:1, and stir well

[0056] (5) Add 35% hot saturated phenol solution to the total volume of the solution in the above solution, then add ultrapure water to make the volu...

Embodiment example 2

[0058] Preparation of high-purity RNA extraction solution:

[0059] The total amount of high-purity RNA extraction solution is 1000ml;

[0060] (1) Preparation of guanidine isothiocyanate solution: take 200ml ultrapure water as a solvent, slowly add 100g of guanidine isothiocyanate, stir evenly on a magnetic stirrer to fully dissolve it;

[0061] (2) Measure the glycerin solution of 50ml, join in the above-mentioned guanidine isothiocyanate solution, carry out sufficient stirring;

[0062] (3) Add 0.8% sodium lauryl sarcosinate relative to the total amount of the solution to the above solution, and stir evenly;

[0063] (4) Add acetic acid solution and sodium acetate solution with a concentration of 0.1M relative to the total amount of the solution, respectively, into the above solution at a volume ratio of 3:1, and stir well;

[0064] (5) Add 40% hot saturated phenol solution to the total volume of the solution in the above solution, then add ultrapure water to make the vol...

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PUM

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Abstract

The invention discloses an RNA extraction solution, a method for preparing the RNA extraction solution, and an RNA reagent extraction method. The RNA extraction solution comprises the mass fraction orthe molar concentration of the following components: 30-50% by volume relative to the total amount of the solution; The total amount of the solution is 3 to 10% by volume of the polyol; the cerium salt is 0.5 to 2 M in concentration relative to the total amount of the solution; 0.1 to 1% by volume of the sodium lauryl sarcosinate relative to the total amount of the solution With respect to the total amount of the solution of 0.05-0.2M concentration of acetic acid; relative to the total amount of the solution 0.05-0.2M of sodium acetate, the remaining component is ultrapure water, in the present invention through acetic acid The ratio of sodium acetate to the pH of the control buffer can completely separate DNA and RNA, and avoid the disintegration of RNA under acidic conditions. It also combines the convenience of silica gel membrane adsorption column and simplifies. The subsequent steps of RNA extraction can be completed by shortening to about 10 minutes compared to the previous onehour or more.

Description

technical field [0001] The invention relates to the biomedicine industry, in particular to an RNA extraction solution, a method for preparing the RNA extraction solution, and an RNA reagent extraction method. Background technique [0002] Ribonucleic acid (abbreviated as RNA, Ribonucleic Acid) is a genetic information carrier that exists in biological cells and some viruses and viroids. RNA is a long chain molecule formed by condensation of ribonucleotides through phosphodiester bonds. RNA is a single strand formed by transcription based on a strand of DNA as a template and based on the principle of complementary base pairing. RNA can regulate the phenotype of organisms through properties such as type and expression level. In order to conduct gene expression analysis and research, it is very important to extract high-purity RNA from various biological samples. Many methods have been developed to obtain RNA from biological materials, such as phenol extraction, precipitation...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1006
Inventor 杨勇
Owner SUZHOU NEW CELL & MOLECULAR BIOTECH CO LTD
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