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Nucleic acid chip, preparation method and application thereof

A nucleic acid chip and nucleic acid technology, applied in biochemical equipment and methods, nucleotide libraries, protein nucleotide libraries, etc., to achieve the effects of low cost, expanded application range, and ease of use

Active Publication Date: 2010-04-21
SUZHOU GENOARRAY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] By adopting surface modification methods and micro-nano technology, the present invention not only solves the problems of traditional reverse transfection chips, but also greatly expands their application scope, such as the screening of functional genes such as cell migration

Method used

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  • Nucleic acid chip, preparation method and application thereof
  • Nucleic acid chip, preparation method and application thereof
  • Nucleic acid chip, preparation method and application thereof

Examples

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Effect test

example 1

[0060] Example 1 cell culture:

[0061] The cells used in the experiment include Hela, HepG2 and U2-OS. The cell culture medium was Dulbecco's Modified Eagle Medium (DMEM, Invitrogen), which contained 10% fetal bovine serum and 1% double antibody. Culture conditions: humidity 95%, temperature 37°C, 5% CO 2 . When the cells are subcultured, wash them with PBS first, then add 0.25% trypsin containing 5mM EDTA, and add serum to terminate the digestion when the edges of the cells are observed to float and appear round. Use a pipette to blow off the adherent cells, centrifuge to remove the supernatant, and obtain the deposited cells. After adding an appropriate amount of medium to dilute, use a cell counting plate to count, and then culture.

example 2

[0062] Example 2 utilizes dispenser to prepare nucleic acid lattice

[0063] Nucleic acid array preparation was performed using a dispenser based on microfluidic technology (Wang, et al. Lap Chip, 2009). According to different experimental requirements, other spotting instruments can also be used. The dispenser is designed with six independent pipes so that six different samples can be dispensed simultaneously. An air pressure pulse (0.04 MPa, 15 msec) generated by compressed air can drive approximately 110 nanoliters of liquid reagent to fill the surface of a well that is not covered by polymer. When spotting, the chip was placed on a 37°C temperature-controlled platform to prevent liquid reagents from dissolving the polymer. After spotting, clean the distributor tube with ultrapure water and dry it with compressed air for next use.

example 3

[0064] Example 3 Cell Self-Assembled Small Islands

[0065] Clean the surface of a glass slide (25 mm X 25 mm) commonly used in the laboratory with detergent and ultrapure water. After drying, drop a layer of polymer film on the surface, use 65 microliters of 6% (w / v) Poly(N-isopropylacrylamide) (Sigma or PolySciences) ethanol solution, dry, and store at room temperature for 12 hours. Prepare a silicon wafer mask according to the experimental requirements, and micro-holes of different sizes or shapes can be engraved on it by using micro-processing technology. Specifically, the silicon wafer mask is covered on the glass slide after the film is dried, and etched in an oxygen plasma etching machine. The etching conditions are 50pa oxygen pressure, 200W power, and the etching time is about 3.5 minutes. After the etching is completed, put the glass slide into the ultra-clean bench for ultraviolet irradiation disinfection, and then use the dispenser to process the siRNA microarray...

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Abstract

The invention relates to a nucleic acid chip and a preparation method and application thereof, belonging to the detection field of enzymes, microorganisms or nucleic acid molecules. The nucleic acid chip comprises a substrate, nucleic acid molecules are attached to a part of the surface of the substrate; a region of the substrate without the nucleic acid molecules is covered with a material capable of inhibiting cell growth; the material can be a non-biocompatible material, such as poly-N-isopropylacrylamide, polyvinyl caprolactam, polyacrylic acid, chitosan, sodium alginate, and the like or be DNA, siRNA, esiRNA, plasmid, and the like. The nucleic acid chip is particularly suitable for researching functional gene screening aspects, such as cell proliferation, cell differentiation, apoptosis, cell migration, and the like.

Description

technical field [0001] The invention relates to the field of detection of enzymes, microorganisms or nucleic acid molecules. Specifically, the present invention relates to a nucleic acid chip that can be used for screening functional genes such as cell migration, cell proliferation, cell differentiation, and apoptosis, as well as its preparation method and application. Background technique [0002] RNA interference (RNA interfering, RNAi) is a phenomenon in which double-stranded RNA (double-stranded RNA, dsRNA) induces efficient and specific degradation of homologous mRNA. It is an ancient and evolutionarily highly conserved gene expression regulation mechanism in the biological world. In 1998, researchers discovered that injecting dsRNA into C.elegans or Drosophila can specifically inhibit gene expression, and called this phenomenon of inhibiting specific gene expression triggered by dsRNA as RNAi. Subsequently, the basic research and application of RNAi quickly became one...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/02C40B40/06C40B50/06
CPCC40B40/08C40B40/04B01J2219/00743B01J2219/00635B01J2219/00637G01N33/552B01J2219/00722B01J19/0046B01J2219/00608
Inventor 席建忠黄岩谊
Owner SUZHOU GENOARRAY
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