Hydrogel-based digital PCR chip, and application method thereof

A hydrogel and chip technology, applied in chemical instruments and methods, biochemical equipment and methods, laboratory containers, etc., can solve the limitations of digital PCR technology development and application, complex digital PCR chip manufacturing process, and operating procedures Complicated and other issues, to achieve the effect of novel design, no cross-contamination, and good biocompatibility

Pending Publication Date: 2018-10-09
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these sample dispersion methods have some similar defects, such as high cost, complicated operating procedures, and the need for additional auxiliary equipment, etc.
For example, in droplet dispersion, dispersing tens of microliters of sample into tens of thousands of microdroplets requires special and expensive auxiliary equipment; Operated by professionals; a commercial digital PCR instrument usually costs millions of RMB
All these reasons limit the further development and application of digital PCR technology to a certain extent.

Method used

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  • Hydrogel-based digital PCR chip, and application method thereof
  • Hydrogel-based digital PCR chip, and application method thereof
  • Hydrogel-based digital PCR chip, and application method thereof

Examples

Experimental program
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Effect test

example 1

[0053] Example 1. Dispersed pigment solution on a hydrogel digital PCR chip

[0054] Step 1: Add 8 microliters of pigment solution dropwise to the sample hole on one side of the PCR chip, and the pigment solution will automatically flow along the flow channel to the array area where the miniature hydrogel array of the hydrogel digital PCR chip is located. The freeze-dried hydrogel has good water absorption, so that the pigment aqueous solution is dispersed into each microcolumn of the porous hydrogel, and plays the role of dispersing the pigment solution (see Figure 5 ).

[0055] Step 2: After step 1), the sealing oil is injected by applying external pressure through the sample hole, and the sealing oil is used to fill the part outside the micro-hydrogel array, and at the same time discharge the micro-hydrogel array in the array area. air to form 11697 mutually independent airtight microsystems with the microgel column as a unit, so that the added 8 microliters of pigment so...

example 2

[0056] Example 2. Disperse nucleic acid solution with hydrogel digital PCR chip and perform PCR amplification

[0057] Step 1: Mix the diluted standard nucleic acid solutions of different concentrations with the PCR master mix (including PCR polymerase, dNTP, PCR probes, primers, etc.) (up to 8 microliters after mixing), and then drop 8 microliters of the mixed solution Add the sample hole on one side of the PCR chip, and the mixed solution will automatically diffuse along the flow channel to the array area where the micro hydrogel array is located. Due to the porous structure of the hydrogel freeze-dried, the solution in the Nucleic acid molecules are absorbed and dispersed into individual micropillars of the porous hydrogel.

[0058]Step 2: inject sealing oil, use the sealing oil to fill the part outside the micro-hydrogel microarray, and discharge the air in the array area where the micro-hydrogel array is located to form 11697 mutually independent PCR micro-reaction system...

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PUM

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Abstract

The invention provides a hydrogel-based digital PCR chip, and an application method thereof. The hydrogel-based digital PCR chip comprises a micro hydrogel array, a polymethyl methacrylate cover plate, a double faced adhesive tape interface layer, and a glass substrate; the micro hydrogel array contains 2300 to 46000 micro gel units; the volume of each freeze-dried micro gel unit ranges from 0.2 to 5 nano-liters. According to the application method, when a nucleic acid sample solution flows through the micro hydrogel array, the nucleic acid sample solution is absorbed, dispersed, and retainedin each of the micro gel units, and sealing oil is introduced into the system for complete isolation of the reaction system. The hydrogel-based digital PCR chip is novel in design, is convenient to prepare, is low in cost, is high in biocompatibility, is convenient in operation, and is wide in suitable range.

Description

technical field [0001] The invention belongs to the technical application field of biochemistry and molecular biology, and specifically relates to a device for dispersing trace amounts of nucleic acid or other biological sample solutions and a use method. Background technique [0002] Deoxyribonucleic acid (DNA) carries biological genetic information and plays an important role in the storage and transmission of genetic information. The in vitro amplification of DNA is very important to improve the specificity and accuracy of DNA detection. The polymerase chain reaction (Polymerase Chain Reaction, PCR) invented by K.B.Mullis in 1983 made it possible to amplify DNA in vitro. PCR technology realizes qualitative analysis of DNA products through agarose gel electrophoresis after amplification. With the increasing demand for quantitative DNA detection, traditional electrophoresis-based PCR is gradually replaced by quantitative PCR analysis, that is, real-time fluorescent quanti...

Claims

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Application Information

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IPC IPC(8): C12M1/00
CPCB01L3/502707B01L7/52Y02A50/30
Inventor 李菲曹雷
Owner XI AN JIAOTONG UNIV
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