Preparation method of micro-fluidic chip for sequencing

A microfluidic chip and sequencing technology, applied to laboratory containers, chemical instruments and methods, and techniques for producing decorative surface effects, etc., can solve the problems of low sequencing throughput and low accuracy, and achieve detection Simplicity, high precision, large effective area effect

Active Publication Date: 2015-07-15
BEIJING ZHONGKEZIXIN TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing microfluidic gene chip still has the disadvantages of low sequencing throughput and low accuracy, so it is necessary to prepare a new type of gene chip to meet the needs of practical applications

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for preparing a microfluidic chip for sequencing, comprising the following steps:

[0023] Step 1: Take a single-sided polished substrate, the chip has a chip length of 49mm, a width of 48mm, a thickness of 2mm, and the material is an optical fiber;

[0024] Step 2: Bath in N,N-dimethylformamide water for 20 minutes at 90°C, then rinse with water for 1 minute; blow dry;

[0025] Step 3: Apply photoresist on the polished surface of the chip, and form a reaction pool pattern on the surface by exposure. The effective area of ​​the chip is 27.6x27.6mm 2 ;

[0026] Step 4: Etch the chip by plasma etching to form 700,000 to 710,000 reaction pools with a diameter of 28 μm;

[0027] Step 5: Bath in N,N-dimethylformamide water for 30 minutes, soak in sulfuric acid hydrogen peroxide mixture for 30 minutes, H 2 SO 4 and H 2 o 2 The volume ratio is 3:1, rinse with water for 1.5min;

[0028] Step 6: Complete the preparation of the chip. Spray boron carbide paint eve...

Embodiment 2

[0030] A method for preparing a microfluidic chip for sequencing, comprising the following steps:

[0031] Step 1: Take a single-sided polished substrate, the chip has a chip length of 49mm, a width of 48mm, a thickness of 2mm, and the material is an optical fiber;

[0032] Step 2: Bath in N,N-dimethylformamide for 30 minutes at 85°C, then rinse with water for 2 minutes; blow dry;

[0033] Step 3: Apply photoresist on the polished surface of the chip, and form a reaction pool pattern on the surface by exposure. The effective area of ​​the chip is 27.6x27.6mm 2 ;

[0034] Step 4: Etching the chip to form 700,000 to 710,000 reaction pools with a diameter of 29 μm;

[0035] Step 5: Bath in N,N-dimethylformamide water for 20 minutes, soak in sulfuric acid hydrogen peroxide mixture for 60 minutes, H 2 SO 4 and H 2 o 2 The volume ratio is 2:1, rinse with water for 1min;

[0036] Step 6: Complete the preparation of the chip.

Embodiment 3

[0038] A method for preparing a microfluidic chip for sequencing, comprising the following steps:

[0039] Step 1: Take a single-sided polished substrate, the chip has a chip length of 49mm, a width of 48mm, a thickness of 2mm, and the material is an optical fiber;

[0040] Step 2: Bath in N,N-dimethylformamide water for 25 minutes at 100°C, then rinse with water for 4 minutes; blow and dry;

[0041] Step 3: Apply photoresist on the polished surface of the chip, and form a reaction pool pattern on the surface by exposure. The effective area of ​​the chip is 27.6x27.6mm 2 ;

[0042] Step 4: Etch the chip by plasma etching to form 690,000 to 700,000 reaction pools with a diameter of 30 μm;

[0043] Step 5: Bath in N,N-dimethylformamide water for 20 minutes, soak in sulfuric acid hydrogen peroxide mixture for 40 minutes, H 2 SO 4 and H 2 o 2 The volume ratio is 3:1, rinse with water for 3 minutes;

[0044] Step 6: Complete the preparation of the chip.

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Abstract

The invention belongs to the technical field of manufacturing of biomedical engineering equipment and related devices, and in particular relates to a preparation method of a micro-fluidic chip for sequencing. The preparation method comprises the following steps: 1. preparing a single-surface polished substrate; 2. carrying out water bath with N,N-dimethyl formamide and flushing with water, blowing the substrate and drying; 3. coating the polished surface of a chip with a photoresist, and exposing to form a reaction tank pattern on the surface; 4. by virtue of dry etching, forming a reaction tank; 5. carrying out water bath with N,N-dimethyl formamide, soaking in a sulfuric acid and hydrogen peroxide mixed solution and flushing with water; and 6. completing the preparation of the chip. According to the preparation method disclosed by the invention, interference of impurities or stains on a sequencing reaction can be avoided, and the sequencing accuracy and the sequencing efficiency can be guaranteed.

Description

technical field [0001] The invention belongs to the field of biomedical engineering equipment manufacturing technology and related devices, and in particular relates to a preparation method of a microfluidic chip for sequencing. Background technique [0002] Because the gene chip technology immobilizes a large number of probes on the support at the same time, it can detect and analyze a large number of sequences in the sample at one time, thereby solving the problem of complicated operation, low degree of automation, small number of operation sequences and detection efficiency of traditional nucleic acid imprinting hybridization technology. Low and insufficient. Moreover, by designing different probe arrays and using specific analysis methods, this technology can have a variety of different application values, such as gene expression profiling, mutation detection, polymorphism analysis, genome library mapping and hybridization sequencing, etc. [0003] The Sanger sequencing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/00B81C1/00
Inventor 陈哲张睿王者馥王绪敏殷金龙任鲁风
Owner BEIJING ZHONGKEZIXIN TECH
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