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Ultrasonic-based nucleic acid extraction method

An extraction method, ultrasonic technology, applied in biochemical equipment and methods, microbial measurement/inspection, DNA preparation, etc., can solve the problems of complex nucleic acid extraction process and low nucleic acid purity, and shorten the homogenization time and elution time Short, good purification effect

Pending Publication Date: 2022-01-04
SUZHOU DIYINAN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current nucleic acid extraction methods still have defects such as complicated nucleic acid extraction process and low nucleic acid purity.

Method used

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  • Ultrasonic-based nucleic acid extraction method

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Experimental program
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Effect test

Embodiment 1

[0058] This embodiment provides a method for nucleic acid extraction based on ultrasound. The whole blood sample is divided into two equal parts, each with 200 μL, and the two identical whole blood samples are subjected to the following nucleic acid extraction:

[0059] (1) Sample lysis: Use a pipette gun to sequentially pipette 200 μL whole blood sample, 500 μL chemical lysate (pH=7.2), 10 μL proteinase K and 20 μL magnetic bead suspension into the cuvette, and sonicate the reaction solution for 1 min at room temperature , the ultrasonic power is 10W, the ultrasonic frequency is 20kHz, and the whole blood sample is lysed under the action of ultrasonic, lysate and proteinase K;

[0060] Wherein, the chemical lysate used comprises 1mol / L sodium acetate, 3mol / L guanidine hydrochloride, 3mol / L EDTA, 0.5vol% sodium dodecylbenzenesulfonate and 50vol% polyethylene glycol;

[0061] (2) Nucleic acid purification: After the sample lysis is completed, the reaction solution in the reacti...

Embodiment 2

[0065] This embodiment provides a method for nucleic acid extraction based on ultrasound. The whole blood sample is divided into two equal parts, each with 200 μL, and the two identical whole blood samples are subjected to the following nucleic acid extraction:

[0066] (1) Sample lysis: Use a pipette gun to sequentially draw 200 μL whole blood sample, 500 μL chemical lysate (pH=7), 10 μL proteinase K and 20 μL magnetic bead suspension into the cuvette, and continuously sonicate the reaction solution at room temperature 2min, the ultrasonic power is 3W, the ultrasonic frequency is 10kHz, and the whole blood sample is lysed under the action of ultrasonic, lysate and proteinase K;

[0067] Wherein, the chemical lysate used comprises 0.5mol / L sodium acetate, 2mol / L guanidine hydrochloride, 1mol / L EDTA, 0.25vol% Triton X-100 and 40vol% polyethylene glycol;

[0068] (2) Nucleic acid purification: After the sample lysis is completed, the reaction solution in the reaction container i...

Embodiment 3

[0072] This embodiment provides a method for nucleic acid extraction based on ultrasound. The whole blood sample is divided into two equal parts, each with 200 μL, and the two identical whole blood samples are subjected to the following nucleic acid extraction:

[0073] (1) Sample lysis: Use a pipette gun to sequentially draw 200 μL whole blood sample, 500 μL chemical lysate (pH=7.1), 10 μL proteinase K and 20 μL magnetic bead suspension into the cuvette, and continuously sonicate the reaction solution at room temperature 3min, the ultrasonic power is 5W, the ultrasonic frequency is 18kHz, and the whole blood sample is lysed under the action of ultrasonic, lysate and proteinase K;

[0074] Wherein, the chemical lysate used comprises 0.7mol / L sodium acetate, 2.5mol / L guanidine hydrochloride, 2mol / L EDTA, 0.3vol% Tween 20 and 45vol% polyethylene glycol;

[0075] (2) Nucleic acid purification: After the sample lysis is completed, the reaction solution in the reaction container is...

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Abstract

The invention provides an ultrasonic-based nucleic acid extraction method which comprises the following steps: putting a sample to be detected, a chemical lysis solution, protease K and a magnetic bead suspension into a reaction container, performing sample lysis under the action of ultrasonic waves, then performing nucleic acid purification, and finally performing nucleic acid elution through ultrasonic treatment and / or high-temperature treatment. According to the nucleic acid extraction method provided by the invention, the ultrasonic method is matched with the chemical lysis solution and the protease K to act on the sample at the same time, so that the lysis efficiency is effectively improved, and the sample lysis can be completed at normal temperature within 30-60s. According to the nucleic acid extraction method provided by the invention, an ultrasonic method is matched with a chemical lysis solution and protease K to act on a sample at the same time, so that the lysis efficiency is effectively improved, and the lysis of samples such as cells and microorganisms can be completed within 30-60s at normal temperature.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid extraction, and relates to an ultrasonic-based nucleic acid extraction method. Background technique [0002] Nucleic acid includes two kinds of molecules, DNA and RNA. Biotechnology with nucleic acid as the research object, including a series of technologies such as extraction, cloning, amplification, detection, and sequencing of nucleic acid, has developed rapidly in recent years. These technologies are currently not only used in research units , and has been widely used in multiple functional departments in social life. The first step in nucleic acid research is to extract nucleic acid from various biological samples, and the efficiency of nucleic acid extraction becomes a restrictive factor for the success of downstream nucleic acid research. Nucleic acid exists in the state of binding to protein in cells. The main steps of nucleic acid extraction are: lysing cells to remove protein bound...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2521/537C12Q2523/301C12Q2523/308C12Q2523/107
Inventor 胡杰巩燕高运通孔梦奇黄培杨仁杰徐峰
Owner SUZHOU DIYINAN BIOTECHNOLOGY CO LTD