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Micro-fluidic chip and nucleic acid extraction and purification method therewith

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

Active Publication Date: 2016-07-06
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Micro-fluidic chip and nucleic acid extraction and purification method therewith
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  • Micro-fluidic chip and nucleic acid extraction and purification method therewith

Examples

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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 superparamagnetic nanoparticles. Under the acidic condition of pH 6.1, the nucleic acid and superparamagnetic nanoparticles combine to form nucleic acid-magnetic nanoparticle complexes and a...

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, and controlled by a pneumatic microvalve, The real sample, superparamagnetic silica beads, loading 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 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 com...

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Abstract

The invention provides a micro-fluidic chip and a nucleic acid extraction and purification method therewith. The micro-fluidic chip has a four-layer membrane structure including: a liquid path layer, a gas path control layer, a magnet control layer and a glass substrate layer. The gas path control layer is arranged below the liquid path layer. The substrate layer is arranged below the gas path control layer. The magnet control layer is arranged below the glass substrate layer. The method, with the micro-fluidic chip, includes the steps of: a) introducing a target sample; b) introducing superpara-magnetic silicon beads for nucleic acid purification; and c) extracting and purifying nucleic acid from the mixed sample. In the invention, introduction of the sample, cleaning of non-nucleic acid substances and collection and purification of the nucleic acid are integrated on one micro-fluidic chip, so that the nucleic acid extraction and purification process is optimized and extraction and purification efficiency is greatly improved. The method simplifies manual operations and achieves temporal-spatial resolution information which is difficult in conventional methods, and has excellent nucleic acid extraction and purification effects.

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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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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