Extract and application thereof in preserving tissue or cell and extracting RNA (Ribonucleic Acid)

A technology for extracting liquid and tissue, applied in the field of extracting liquid, can solve the problems of incompatibility and low efficiency, and achieve the effects of stable properties, high purity, and simplified operation steps

Active Publication Date: 2019-04-09
CHENGDU DAOSHENG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem of low efficiency caused by the incompatibility between the existing solution preservation and the existing extraction solution in the prior art, the present invention provides an extraction solution and the extraction solution when preserving tissues or cells, extracting Applications in RNA

Method used

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  • Extract and application thereof in preserving tissue or cell and extracting RNA (Ribonucleic Acid)
  • Extract and application thereof in preserving tissue or cell and extracting RNA (Ribonucleic Acid)

Examples

Experimental program
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Embodiment 1

[0061] This embodiment provides an extraction solution, including 1.6 mol / L guanidine thiocyanate and 1 mol / L guanidine hydrochloride, and the pH of the extraction solution is adjusted to 4 by acetic acid. The extract is applied to the preserved tissue or cells.

Embodiment 2

[0063] The present embodiment provides an extract, comprising:

[0064]

[0065] Wherein, the pH of the extract is adjusted to 4 by acetic acid. This extract solution was used to extract RNA.

Embodiment 3

[0067] This embodiment provides an extract, including extract A and extract B;

[0068] Wherein extract A comprises the guanidine thiocyanate of 1.6mol / L and the guanidine hydrochloride of 1mol / L;

[0069] The extract B includes 57vt% water-saturated phenol, 0.24mol / L guanidine thiocyanate, 0.6mol / L ammonium thiocyanate, 11.25vt% glycerin and 0.01mol / L SDS; the balance is solvent.

[0070] The pH of the extract A and the extract B are both 4.

[0071] The amount of RNA extracted by using the extract solution in this example is 1000-1800 μg / mL, the measured A260 / A280 is in the range of 1.6-1.9, and the A260 / A230 is in the range of 1.6-1.9. Agarose gel electrophoresis pictures such as figure 1 shown.

[0072] Adopt above-mentioned extraction solution A and extraction solution B to carry out RNA extraction method, concrete steps are as follows:

[0073] 1. Tissues or cells preserved with extract A, said extract A includes 1.2-2.0 mol / L guanidine thiocyanate and 0.5-1.5 mol / L ...

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Abstract

The invention discloses an extract which comprises an extract A and an extract B, wherein the extract A applied to tissue or cell preservation; the extract A comprises 1.2-2.0mol/L guanidine thiocyanate and 0.5-1.5mol/L guanidine hydrochloride; the extract B comprises 30-65vt% of water saturated phenol, 9-15vt% of glycerinum and the balance of a solvent in percentage by volume; the extract B further comprises 0.20-1mol/L guanidine thiocyanate, 0.4-0.8mol/L ammonium thiocyanate and a 0.0073-0.018mol/L surfactant by mole concentrations; the pH values of both the extract A and the extract B are 3.5-4.5; the extract B is applied to RNA extraction. Ratios of components of the extract disclosed by the invention are reasonably designed, the components have mutual synergistic effects, and extracted RNA is high in purity, high in yield and low in protein contamination.

Description

technical field [0001] The invention belongs to the technical field of biological preparations, and in particular relates to an extract and the application of the extract in preserving tissues or cells and extracting RNA. Background technique [0002] In modern molecular biology experiments and clinical molecular diagnosis, it is often necessary to isolate and purify RNA from tissues and cells, and the quality of RNA often affects the success or failure of molecular biology experiments such as cDNA library, RT-PCR and Northern Blot. [0003] The commonly used method is to first protect the tissue with a tissue protection solution, and then use the extraction solution to extract the RNA in the tissue or cells. Commonly used protection methods include liquid nitrogen protection and protection solution protection, and the commonly used protection solutions include RNA later reagents and RNA safety reagents. [0004] Among them, the liquid nitrogen protection method not only re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 赵伟廖东升赵仲玖邱坤
Owner CHENGDU DAOSHENG BIOTECH CO LTD
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