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Extract and application thereof in RNA extraction

A technology of extraction solution and molar concentration, which is applied in the extraction of RNA. In the field of extraction solution, it can solve the problems of incompatibility and low efficiency, and achieve the effect of stable properties, high purity and reasonable design.

Active Publication Date: 2019-04-12
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 application of the extraction solution in RNA extraction

Method used

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  • Extract and application thereof in RNA extraction
  • Extract and application thereof in RNA extraction

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Embodiment 1

[0051]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

[0053] The present embodiment provides an extract, comprising:

[0054]

[0055] Wherein, the pH of the extract is adjusted to 4 by acetic acid. This extract solution was used to extract RNA.

Embodiment 3

[0057] This embodiment provides an extract, including extract A and extract B;

[0058] Wherein extract A comprises the guanidine thiocyanate of 1.6mol / L and the guanidine hydrochloride of 1mol / L;

[0059] 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.

[0060] The pH of the extract A and the extract B are both 4.

[0061] 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.

[0062] Adopt above-mentioned extraction solution A and extraction solution B to carry out RNA extraction method, concrete steps are as follows:

[0063] 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 present invention discloses an extract and application thereof in RNA extraction. The extract includes, in terms of volume fraction, 30-65 vt% of water-saturated phenol, 9-15 vt% of glycerol and the balance of a solvent; an extract B also includes, in terms of molar concentration, 0.20-1 mol / L of guanidine thiocyanate, 0.4-0.8 mol / L of ammonium thiocyanate and 0.0073-0.018 mol / L of a surfactant; and the pH of the extract is 3.5-4.5. Components in the extract of the invention are reasonable in proportion design and have synergistic effects with one another, so that the purity of the extracted RNA is high, the yield is high, and the protein pollution is low; the extract are stable in performance, not easily affected by season, humidity and other conditions, and can be fully used in combination, so that the purpose of rapid, low-cost, high-quality and large-scale extraction of animal RNA in the laboratory can be realized.

Description

technical field [0001] The invention belongs to the technical field of biological preparations, and in particular relates to an extraction solution and the application of the extraction solution in RNA extraction. 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 RNAlater reagent and RNAsafety reagent. [0004] Among them, the liquid nitrogen protection method not only requires harsh...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 邱坤赵伟廖东升赵仲玖
Owner CHENGDU DAOSHENG BIOTECH CO LTD
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