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Method for extracting ribonucleic acid from blood

A ribonucleic acid and blood technology, applied in the field of nucleic acid purification in biology, can solve the problems of high cost and low process efficiency, and achieve the effects of good integrity, high purity and efficient extraction process

Inactive Publication Date: 2014-08-06
谭晓刚
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a new method for rapidly extracting ribonucleic acid from blood in order to solve the above-mentioned problems in the prior art, so as to solve the technical problems of low efficiency and high cost of the existing process

Method used

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  • Method for extracting ribonucleic acid from blood
  • Method for extracting ribonucleic acid from blood
  • Method for extracting ribonucleic acid from blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Add 400 microliters of rapid cell lysis solution to 100 microliters of human whole blood sample, the lysis solution contains 0.5% by weight and volume of dodecyltrimethylammonium chloride, and mix the mixture thoroughly , And let the mixture stand for 20 minutes at room temperature to form a precipitate of dodecyltrimethylammonium chloride and ribonucleic acid;

[0038] (2) Centrifuge the sediment in (1) with a centrifugal force of 7000g and a centrifugal time of 8 minutes to remove the upper layer solution after centrifugation and retain the sediment;

[0039] (3) Add 500 microliters of nuclease-free water to the precipitate in (2), and mix thoroughly; among them, because nuclease will destroy ribonucleic acid, the reagent for cleaning the precipitate cannot contain nuclease, which can be used in the field Commonly used cleaning agents, or pure water, the purpose of cleaning is to remove other impurities in the sediment, such as proteins;

[0040] (4) Centrifuge the sedi...

Embodiment 2

[0046] (1) Add 300 microliters of fast cell lysate to 100 microliters of mouse whole blood sample, the lysate contains 0.2% tetradecyltrimethylammonium chloride and 5% Tween by weight and volume. -20. Mix the mixture thoroughly, and let the mixture stand for 15 minutes at room temperature to form a precipitate of tetradecyltrimethylammonium chloride and ribonucleic acid;

[0047] (2) Centrifuge the sediment in (1) with a centrifugal force of 5000g and a centrifugal time of 10 minutes to remove the upper layer solution after centrifugation and retain the sediment;

[0048] (3) Add 400 microliters of nuclease-free water to the precipitate in (2) and mix well;

[0049] (4) Centrifuge the sediment in (3) with a centrifugal force of 5000 g and a centrifugal time of 10 minutes to remove the upper layer solution after centrifugation and retain the sediment;

[0050] (5) Add 50 microliters of ribonucleic acid purification solution to the precipitate in (4). The purified solution contains 6% b...

Embodiment 3

[0053] (1) Add 500 microliters of fast cell lysate to 100 microliters of human whole blood sample, the lysate contains 0.7% cetyltrimethylammonium bromide and 10% TritonX- 100. Mix the mixture thoroughly, and let the mixture stand for 30 minutes at room temperature to form a precipitate of cetyltrimethylammonium bromide and ribonucleic acid;

[0054] (2) Centrifuge the sediment in (1) with a centrifugal force of 10000g and a centrifugal time of 5 minutes to remove the upper layer solution after centrifugation and retain the sediment;

[0055] (3) Add 600 microliters of nuclease-free water to the precipitate in (2) and mix well;

[0056] (4) Centrifuge the sediment in (3) with a centrifugal force of 10000g and a centrifugal time of 5 minutes to remove the upper layer solution after centrifugation and retain the sediment;

[0057] Repeat (3) and (4) twice, until the precipitate does not show obvious red;

[0058] (5) Add 150 microliters of ribonucleic acid purification solution to the pr...

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Abstract

The invention discloses a method for extracting ribonucleic acid from blood. The method comprises the steps of adding a rapid cell lysis solution containing a cationic surfactant into whole blood sample, mixing uniformly and standing; carrying out a centrifugal separation after cleaning to remove a supernatant solution and retain precipitate; adding a high-ion-concentration purified ribonucleic acid solution to the precipitate, mixing uniformly and standing; carrying out a centrifugal separation, taking the centrifugally separated solution portion to obtain a ribonucleic acid solution. The method for extracting ribonucleic acid from blood disclosed by the invention is more efficient, rapid and simple and the separation of white blood cells is not carried out firstly. Compared with the prior art, ribonucleic acid with better integrity and higher purity can be obtained by the extraction method and the extraction method is more suitable for experiments, research and analysis on ribonucleic acid and related technologies.

Description

Technical field [0001] The invention relates to the technical field of nucleic acid purification in biology, in particular to a method for rapidly extracting ribonucleic acid from blood. Background technique [0002] Blood is the liquid tissue in the circulatory system of humans and animals. It maintains the connection between the entire body and the external environment, as well as the interconnection between various organs and tissues, and plays an important role in maintaining life. Because it is relatively easy to obtain, it has a very important position in basic and clinical medical research. Blood can be used as a molecular marker in the study of hematopoietic system diseases and in the development of in vitro diagnostic systems, and can also be used for molecular monitoring of many system diseases outside the blood system. [0003] Among them, the effective extraction of ribonucleic acid (RNA for short) in blood is the most important process. Due to the coagulability of bl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 谭晓刚
Owner 谭晓刚
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