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A microfluidic chip and nucleic acid extraction and purification method using the microfluidic chip

A technology of microfluidic chip and purification method, which is applied in the field of microfluidic chip and nucleic acid extraction and purification using microfluidic chip, which can solve the problems of environmental pollution, time-consuming and cumbersome, inflexible operation, etc., and achieve the realization of time-space resolution information , Avoid manual operation, optimize the effect of the purification process

Active Publication Date: 2018-06-19
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the extraction methods commonly used in laboratories have many disadvantages: such as phenol / chloroform extraction method and spin column method, which require multiple centrifuges, have relatively high requirements for external equipment, and are easy to cause sample loss; and the use of organic solvents will also cause environmental pollution. Pollution, endangering the health of the operator; time-consuming and cumbersome extraction steps also reduce the speed of nucleic acid detection
The efficiency of nucleic acid purification depends on the flow rate of the purification reagent, the operation is not flexible, and the simultaneous purification of multiple samples cannot be realized, which does not meet the requirements of automatic control

Method used

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  • A microfluidic chip and nucleic acid extraction and purification method using the microfluidic chip
  • A microfluidic chip and nucleic acid extraction and purification method using the microfluidic chip
  • A microfluidic chip and nucleic acid extraction and purification method using the microfluidic chip

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Experimental program
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Effect test

Embodiment 1

[0064] Extraction and purification of lambda DNA

[0065] separately to figure 1 Add 6ng / μL λDNA solution into the sample pool 4 of the chip shown, and add superparamagnetic silicon beads, loading buffer, washing solution and eluent into inlet 1. Under the control of the pneumatic microvalve, the λDNA and superparamagnetic silicon beads The sample buffer, cleaning solution and eluent enter the mixing unit under the condition of vacuum driving force, and the pneumatic microvalve is also controlled to realize the sufficient mixing and purification of the nucleic acid extraction buffer and λDNA, and the sample buffer, cleaning solution and washing solution are collected respectively. dehydration.

[0066] Specific steps include:

[0067] 1. Fully mix the sample solution containing λDNA with the superparamagnetic nanoparticles. Under the acidic condition of pH 6.1, the nucleic acid and the superparamagnetic nanoparticles combine to form nucleic acid-magnetic nanoparticle complex...

Embodiment 2

[0070] Extraction and purification of real sample DNA

[0071] separately to figure 1 Cell lysates (eukaryotic cells or prokaryotic cells) are respectively added to the sample pool 4 of the chip shown, and superparamagnetic silica beads, sample loading buffer, washing solution and eluent are added to the inlet 1. Under the control of the pneumatic microvalve, The real sample, superparamagnetic silica beads, loading buffer, washing solution and eluent enter the mixing unit under the condition of vacuum driving force, and the pneumatic microvalve is also controlled to realize the full mixing of nucleic acid extraction buffer and lysed cell buffer. Purify, and finally collect the eluate containing the purified DNA sample. The specific steps include: 1. Fully mixing the lysed cell solution with the superparamagnetic nanoparticles, and under the acidic condition of pH 6.1, the nucleic acid and the superparamagnetic nanoparticles combine to form a nucleic acid-magnetic nanoparticle...

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Abstract

The invention provides a microfluidic chip and a nucleic acid extraction and purification method using the microfluidic chip. The microfluidic chip has a 4-layer film structure, which is composed of four parts: a liquid path layer, a gas path control layer, a magnet control layer, and a glass substrate layer; the gas control layer is located below the liquid path layer, and the substrate layer is located under the Below the gas control layer; the magnet control layer is located below the base layer; the method uses the above-mentioned microfluidic chip to perform the following steps: A, introducing the target sample; B, introducing superparamagnetic silicon beads for nucleic acid purification; C, Extract and purify nucleic acids from mixed samples. The present invention concentrates the entry of samples, the cleaning of non-nucleic acid substances, and the collection and purification of nucleic acids on a microfluidic chip, optimizes the process of nucleic acid extraction and purification, greatly improves the efficiency of extraction and purification, simplifies human operation procedures, and realizes The spatio-temporal resolution information that is difficult to achieve by traditional methods has been obtained, and better nucleic acid extraction and purification results have been achieved.

Description

technical field [0001] The invention relates to the field of nucleic acid extraction and purification, in particular to a microfluidic chip and a nucleic acid extraction and purification method using the microfluidic chip. Background technique [0002] The separation and purification technology of biomacromolecular ribonucleic acid is the basis for all aspects of molecular biology research and a key technology in life science research and application. The efficiency and quality of nucleic acid sample preparation will directly affect the results of subsequent experiments. At present, the extraction methods commonly used in laboratories have many disadvantages: such as phenol / chloroform extraction method and spin column method, which require multiple centrifuges, have relatively high requirements for external equipment, and are easy to cause sample loss; and the use of organic solvents will also cause environmental pollution. Contamination endangers the health of the operator...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12N15/10
Inventor 秦建华苏文涛姜雷李艳峰
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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