KR101763515B1 - Method and apparatus for detecting dna using graphene/silicon bio-sensor - Google Patents

Method and apparatus for detecting dna using graphene/silicon bio-sensor Download PDF

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KR101763515B1
KR101763515B1 KR1020160011499A KR20160011499A KR101763515B1 KR 101763515 B1 KR101763515 B1 KR 101763515B1 KR 1020160011499 A KR1020160011499 A KR 1020160011499A KR 20160011499 A KR20160011499 A KR 20160011499A KR 101763515 B1 KR101763515 B1 KR 101763515B1
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최석호
김정길
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경희대학교 산학협력단
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    • C12Q2565/00—Nucleic acid analysis characterised by mode or means of detection
    • C12Q2565/60—Detection means characterised by use of a special device
    • C12Q2565/607—Detection means characterised by use of a special device being a sensor, e.g. electrode

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Abstract

The present invention relates to an apparatus for detecting DNA and a method thereof and, more specifically, to an apparatus for detecting DNA using a graphene/silicon bio-sensor and a method thereof, wherein the graphene/silicon bio-sensor having a substrate, a plurality of silicon nano-structures coated with a functional material, a graphene layer, and an electrode is used to identify DNA in accordance with attachment of target DNA forming a pair with respect to a predetermined probe DNA, thereby detecting and sensing precise DNA with high coupling density.

Description

TECHNICAL FIELD [0001] The present invention relates to a DNA detection apparatus using a graphene / silicon biosensor,

The present invention relates to an apparatus and method for detecting DNA using a graphene / silicon biosensor, and more particularly to a method and apparatus for detecting DNA using graphene / silicon biosensor, The present invention relates to a DNS detection apparatus and method for identifying a DNA according to whether a target DNA pair is paired with a predetermined probe DNA using a biosensor.

In recent years, nanostructures have been of great interest in fundamental scientific research as well as the potential for industrial applications. In particular, vertically aligned silicon nanostructures are considered to be ideal nano-based materials as next generation devices that perform functions such as condensing, power generation, energy storage, and sensors because of their high volume-to-area ratio obtained from their vertical structure.

In order to realize the physico-chemical properties required for practical application as a future device using a silicon nanostructure, it is necessary to realize smooth electrical contact between the vertically aligned nanostructures and the electrode.

Conventional biosensors use a method of detecting DNA using a single silicon nanostructure, or spotting a single or double stranded DNA prepared in advance on a predetermined region of a substrate.

However, in the conventional biosensor, there is a high possibility that the attached biomolecules are randomly arranged on the substrate, and the density of the attached region is low.

In addition, the conventional biosensor has a limitation in that the reliability of DNA detection to be measured is low and the reactivity thereof is very low, such as several nA, by using a single silicon nanostructure.

Korean Patent Laid-Open Publication No. 2015-0017422 (Feb. 20, 2017), "Graphene / Silicon Nanowire Molecule Sensor or its Manufacturing Method and Molecular Identification Method Using the Same & Korean Patent No. 10-1050468 (July 13, 2011), "Biochip and Biomolecule Detection System Using It"

The present invention relates to a DNA detection method using a graphene / silicon biosensor capable of detecting DNA having strong reactivity by vertically bonding a graphene layer having high electrical conductivity and flexibility while being able to contact a plurality of uniformly aligned silicon nanostructures, Apparatus and method therefor.

The present invention also relates to a method of coating a functional material containing at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde on a silicon nanostructure, And a DNA detection apparatus and method using the pin / silicon biosensor.

The present invention also provides a DNA detection apparatus and method using a graphene / silicon biosensor capable of precisely detecting and detecting DNA with high binding density by attaching probe DNA to a silicon nanostructure coated with a functional substance .

The present invention also provides a graphene / silicon biosensor comprising a silicon nanostructure having a probe DNA attached thereto and detecting a change in an amount of current depending on whether the probe DNA is attached to a target DNA pairing with the probe DNA, / DNA detection apparatus using silicon biosensor and method thereof.

 A graphene / silicon biosensor according to an embodiment of the present invention includes a substrate, at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde A plurality of silicon nanostructures on which the functional material including the functional material is coated, a graphene layer disposed on the silicon nanostructure, and an electrode formed on the bottom of the substrate and the graphene layer, respectively, DNA is identified according to whether or not target DNA (target DNA) forming a pair is attached to a predetermined probe DNA attached to the surface of the nanostructure.

The silicon nanostructure may be coated with the functional material for binding force of the probe DNA and the functional material may form a strong covalent bond between the surfaces of the silicon nanostructure so that the surface of the silicon nanostructure and the probe DNA Or < / RTI >

The electrode may be formed of at least one of silver (Ag), gold (Au), copper (Cu), aluminum (Al), platinum (Pt), and alloys thereof.

A DNA detection apparatus using a graphene / silicon biosensor according to an embodiment of the present invention includes a substrate, at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde A plurality of silicon nanostructures coated with a functional material containing one of them, a graphene layer disposed on the silicon nanostructure, and an electrode formed on the bottom of the substrate and on the top of the graphene layer, respectively, And a detector for identifying the DNA according to whether or not a target DNA pair forming a pair with the predetermined probe DNA attached to the surface of the silicon nanostructure through the functional material is attached.

The detection unit may detect the DNA by detecting a change in the amount of current depending on whether the probe DNA and the target DNA are attached from the electrode included in the graphen / silicon biosensor.

According to the embodiment of the present invention, DNA having strong reactivity can be detected by vertically bonding a graphene layer having high electrical conductivity and flexibility while being able to contact with a plurality of uniformly aligned silicon nanostructures.

According to an embodiment of the present invention, a functional material including at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde is coated on a silicon nanostructure, .

Further, according to the embodiment of the present invention, by attaching the probe DNA to the silicon nanostructure coated with the functional material, the binding density can be high, and accurate DNA detection and sensing can be performed.

Also, according to the embodiment of the present invention, a change in the amount of current depending on whether or not the probe DNA is attached to the target DNA pairing with the probe DNA is detected in a graphene / silicon biosensor including the silicon nanostructure having the probe DNA attached thereto, can do.

1 shows an example of a graphene / silicon biosensor according to an embodiment of the present invention.
2A to 2D are schematic views of a method of manufacturing a graphene / silicon biosensor according to an embodiment of the present invention.
FIG. 3 illustrates a functional material attached to a surface of a silicon nanostructure of a graphene / silicon biosensor according to an embodiment of the present invention.
4A to 4C show examples of the mechanism of a graphene / silicon biosensor according to an embodiment of the present invention.
5 is a block diagram of a DNA detection apparatus using a graphene / silicon biosensor according to an embodiment of the present invention.
FIGS. 6A to 6D show scanning electron microscopy (SEM) images of a graphene / silicon biosensor according to an embodiment of the present invention.
FIG. 7A is a graph showing the current change curve with respect to the concentration of the probe DNA according to the embodiment of the present invention, and FIG. 7B is a graph showing the current change according to the attachment of the target DNA and the dummy DNA to the probe DNA according to the embodiment of the present invention Curve and response diagrams.
FIG. 8 is a graph showing a result of a recycling characteristic of a graphene / silicon biosensor according to an embodiment of the present invention.
FIGS. 9A to 9C illustrate fluorescence characteristic images according to the adhesion of DNA applied to a graphene / silicon biosensor according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings and accompanying drawings, but the present invention is not limited to or limited by the embodiments.

The terminology used herein is for the purpose of illustrating embodiments and is not intended to be limiting of the present invention. In the present specification, the singular form includes plural forms unless otherwise specified in the specification. It is noted that the terms "comprises" and / or "comprising" used in the specification are intended to be inclusive in a manner similar to the components, steps, operations, and / Or additions.

As used herein, the terms "embodiment," "example," "side," "example," and the like should be construed as advantageous or advantageous over any other aspect or design It does not.

Also, the term 'or' implies an inclusive or 'inclusive' rather than an exclusive or 'exclusive'. That is, unless expressly stated otherwise or clear from the context, the expression 'x uses a or b' means any of the natural inclusive permutations.

Also, the phrase "a" or "an ", as used in the specification and claims, unless the context clearly dictates otherwise, or to the singular form, .

Furthermore, the terms first, second, etc. used in the specification and claims may be used to describe various elements, but the elements should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another.

Unless defined otherwise, all terms (including technical and scientific terms) used herein may be used in a sense commonly understood by one of ordinary skill in the art to which this invention belongs. Also, commonly used predefined terms are not ideally or excessively interpreted unless explicitly defined otherwise.

In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear. The terminology used herein is a term used for appropriately expressing an embodiment of the present invention, which may vary depending on the user, the intent of the operator, or the practice of the field to which the present invention belongs. Therefore, the definitions of these terms should be based on the contents throughout this specification.

1 shows an example of a graphene / silicon biosensor according to an embodiment of the present invention.

The graphene / silicon biosensor 100 according to an exemplary embodiment of the present invention may be configured such that a predetermined DNA probe attached to a surface of a plurality of silicon nanostructures coated with a functional material Lt; / RTI >

The graphene / silicon biosensor 100 according to an embodiment of the present invention includes a substrate 110, a silicon nanostructure 120, a graphene layer 130, and an electrode 140.

The substrate 110 may be a silicon (Si) substrate.

The silicon nanostructure 120 is aligned and bonded to the substrate 110 and includes at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde for improving the adhesion of DNA Are coated, and a plurality of functional materials are formed.

The silicon nanostructure 120 may be integrally coupled to the substrate 110.

Also, the plurality of silicon nanostructures 120 may be arranged at regular intervals, and the intervals and heights may be arranged at a constant height and spacing. In some embodiments, the plurality of silicon nanostructures 120 may be silicon Or may be a structure in which the head portion of the nanostructure 120 is bundled to form a bundle.

According to an embodiment, the silicon nanostructures 120 disposed on the substrate 110 may be vertically aligned, but the present invention is not limited to the perpendicular angle of 90 degrees, and may be inclined at an oblique angle have.

The silicon nanostructure 120 may include a silicon nanostructure 120 in which an impurity is not present, but may be a p-type or n-type silicon nanostructure. The p-type silicon nanostructure may be doped with a p-type impurity, and the n-type silicon nanostructure may be doped with an n-type impurity.

The n-type impurity may include a Group 5 chemical element such as phosphorus (P) and arsenic (As), and the p-type impurity may include a Group 3 chemical element such as boron (B) can do.

The graphene layer 130 is disposed on the silicon nanostructure 120.

The graphene layer 130 may be a conductive material having a thickness of one layer of atoms with the carbon atoms forming a honeycomb arrangement in a two-dimensional fashion. In addition, the graphene layer 130 is structurally and chemically very stable, and is an excellent conductor, having a higher charge mobility than silicon and capable of flowing more current than copper.

Depending on the embodiment, the graphene layer 130 may be a single-layer graphene in the form of a thin film or a two-layer graphene, but is not limited thereto.

Electrodes 140 are formed on the bottom of the substrate 110 and on the top of the graphene layer 130, respectively.

For example, the electrode 140 may be a metal electrode, and the metal may include silver (Ag), gold (Au), copper (Cu), aluminum (Al), platinum have.

The graphene / silicon biosensor 100 according to an exemplary embodiment of the present invention may be configured such that DNA is attached to a predetermined probe DNA attached to the surface of the silicon nanostructure 120 through a functional material, .

The probe DNA may be a single stranded DNA containing a nucleotide sequence in a sample DNA extracted from a sample cell, and the target DNA may be a single stranded DNA paired with a nucleotide sequence in the sample DNA .

In addition, the graphene / silicon biosensor 100 according to an embodiment of the present invention may exhibit Schottky diode characteristics, and the graphene / silicon biosensor 100 having a Schottky diode characteristic may be a general diode And the threshold voltage is relatively low. As a result, since the efficiency of the circuit is high, the distortion of the signal is small and the measurement efficiency as a biosensor for identifying the DNA can be enhanced.

Hereinafter, the manufacturing process of the graphene / silicon biosensor 100 will be described in detail with reference to FIGS. 2A to 2D.

2A to 2D are schematic views of a method of manufacturing a graphene / silicon biosensor according to an embodiment of the present invention.

Referring to FIG. 2A, a method of fabricating a graphene / silicon biosensor according to an embodiment of the present invention includes forming a substrate 110 on which a gold mesh 111 is deposited. The substrate 110 may be a silicon substrate.

Referring to FIG. 2B, a method of fabricating a graphene / silicon biosensor according to an embodiment of the present invention includes forming a plurality of silicon nanostructures 120 from a substrate 110 using an electrochemical etching method.

2B, an electrochemical etching method for fabricating a graphene / silicon biosensor according to an embodiment of the present invention includes a step of forming a mixed solution of silver nitrate (AgNO 3 ) and hydrofluoric acid (HF) And coating the silver particle on the silicon particle surface, and treating the silver particle coated substrate 110 with deionized water to remove the mixed solution remaining on the substrate 110. [

In addition, the substrate 110 coated with silver particles may be immersed in a mixed solution of hydrofluoric acid, hydrogen peroxide (H 2 O 2 ), and deionized water.

When the substrate 110 is treated with a silver nitrate and hydrofluoric acid mixture solution, phosphorus metal can be used as a catalyst in the silicon etching. The silver nitrate can be mixed in the concentration of 0.001 to 0.05 M and the hydrofluoric acid can be mixed in the range of 1 to 10 M .

The silver particles placed on the silicon have a high electric affinity property, so they take the electrons from the silicon surface in contact with the silver particles rather than the silicon surface exposed to the solution, and the hydrogen peroxide in the solution is reduced to water by the electrons, .

At the same time as the electrons are taken away, the silicon that acquires the holes is oxidized and the oxidized silicon can be removed by hydrofluoric acid. In this continuous reaction, the thin film coated with the silver particles is etched with silicon and the silicon nanostructure 120 is formed between the silver foil membrane holes.

For example, when the mixed solution having a volume ratio of HF / H 2 O 2 / H 2 O of 1 / 0.2 / 2 to the silicon nanostructure 120 is etched for 10 minutes at room temperature, the silicon nanostructure 120, The length of the silicon nanostructure 120 may be about 17 mu m, and the head portion of the silicon nanostructure 120 may be bundled.

However, if the volume ratio of HF / H 2 O 2 / H 2 O is varied, the length of the final silicon nitride structure 120 can be controlled and the porosity can be controlled according to the concentration of hydrogen peroxide And the arrangement of the silicon nanostructures 120 can be controlled by adjusting the number of the porous structures.

2B, a method of manufacturing a graphene / silicon biosensor according to an embodiment of the present invention includes the steps of forming 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde (Glutaraldehyde) may be coated on the functional material.

The functional material may be a material that improves adhesion so that the DNA is well adhered to the surface of the silicon nanostructure 120. The method of manufacturing the graphene / (3-APTES) and glutaraldehyde may be coated on the surface of the silicon nanostructure 120. The functional nano-structure of the silicon nanostructure 120 may be coated with the functional material.

Referring again to FIG. 2C, a method of fabricating a graphene / silicon biosensor according to an embodiment of the present invention includes joining a graphene layer 130 onto a silicon nanostructure 120 coated with a functional material.

The graphene layer 130 can be grown using chemical vapor deposition (CVD), and the graphene layer 130 grown to a large area can be transferred onto the silicon nanostructure 120.

More specifically, in the preparation of graphene by chemical vapor deposition, copper (or nickel) to be used as a catalyst layer is deposited on a substrate and reacted with a mixed gas of methane and hydrogen at a high temperature to deposit an appropriate amount of carbon in the catalyst layer Or adsorbed on the surface of the catalyst layer, and the carbon atoms contained in the catalyst layer are crystallized on the surface to form a graphene crystal structure on the metal.

Thereafter, the catalyst layer is removed from the synthesized graphene thin film to separate graphene from the substrate.

Then, PMMA (polymethylmethacrylate) mixed with polymethyl methacrylate (PMMA) and benzene was spin-coated on the synthesized graphene, and PMMA was coated through the coating with ammonium persulfate ) Solution can be used to hold the graphene and fix it when removing the copper foil.

Thereafter, the copper foil is removed from the ammonium persulfate solution, the ammonium persulfate solution remaining on the graphene is washed with DI water, the washed graphene is transferred onto the silicon nano structure 120, (130) can be formed.

Next, the bonding force between the silicon nanostructure 120 and the graphene layer 130 can be increased through the heat treatment after transferring the graphene layer 130 to the silicon nanostructure 120. After the heat treatment, acetone is used to remove the PMMA present on the graphene, and the PMMA residue remaining on the graphene surface may be removed by heat treatment by rapid thermal annealing to finally form the graphene layer 130.

Referring to FIG. 2D, a method of fabricating a graphene / silicon biosensor according to an embodiment of the present invention includes forming electrodes 140b and 140a on the lower portion of the substrate 110 and on the upper portion of the graphene layer 130, respectively.

The electrode 140 may be a metal electrode and may include silver (Ag), gold (Au), copper (Cu), aluminum (Al), platinum (Pt), or an alloy thereof.

A thermal evaporation method, an electron beam evaporation method, a sputtering evaporation method, or the like may be used to deposit the electrode 140 on the silicon nanostructure 120 transferred with the graphene layer 130. However, the present invention is not limited thereto.

A specific embodiment of manufacturing a graphene / silicon biosensor according to an embodiment of the present invention is as follows.

≪ Fabrication of silicon nano structure >

The silicon to be used for etching was a p-type silicon wafer which was doped with boron and had a resistance of 1-10 Ohm / cm, and the organic substances on the surface of the silicon wafer were removed by a 3: 1 mixture of sulfuric acid and hydrogen peroxide And washed with deionized water.

As a first step to fabricate silicon nanostructures, silicon wafers were immediately mixed with a 0.005M solution of AgNO 3 and 5M hydrofluoric acid for 1 minute at a slow rate to form phosphorous particles with an etch catalyst on the silicon surface Air atmosphere. The remaining solution on the prepared silicon wafer was sufficiently diluted with deionized water and removed.

In the second step, the silicon wafer with the silver particles on the surface was immersed in a mixture of hydrofluoric acid, hydrogen peroxide and deionized water and etched at room temperature for 10 minutes. The concentration of the etching solution was adjusted so that the volume ratios of HF / H 2 O 2 / H 2 O were 1 / 0.2 / 2, 1 / 0.5 / 2, 1 / 0.75 / 2 and 1/1/2.

< Graphene  Production>

Generally, a large area of graphene was produced by using a well-known chemical vapor deposition (CVD) method. First, copper to be used as a catalyst layer is deposited on the substrate, and a methane-hydrogen mixed gas is reacted at a high temperature of about 1000 ° C. so that an appropriate amount of carbon is dissolved or adsorbed in the catalyst layer. After cooling, the carbon atoms contained in the catalyst layer are crystallized on the surface to form a graphene crystal structure.

The thus-synthesized graphene can be separated from the substrate by removing the catalyst layer, and then used for a desired application. Detailed manufacturing and transcription processes are described in previous studies. [J. Appl. Phys. 113, 064305 (2013).

&Lt; Silicon nanostructure - Graphene layer  Fabrication of bonding structure>

A large area of graphene fabricated by chemical vapor deposition (CVD) was supported on PMMA, placed in deionized water, and transferred onto a silicon nanostructure. The transferred sample on the silicon nanostructure was placed on a hot plate, dried at room temperature for 1 hour or more, and further dried at 60 to 100 degrees for 3 hours or more.

 &Lt; Silicon nanostructure - Graphene layer  Fabrication of electrode with bonded structure &gt;

Electrodes (Au, Ag, Pt, etc.) were deposited on the graphene layer by thermal evaporation, electron beam evaporation, sputtering, or the like on the dried silicon nano structure-graphene layer. In one embodiment of the present invention, gold (Au) is deposited on the top and silver (Ag) is deposited on the bottom electrode.

FIG. 3 illustrates a functional material attached to a surface of a silicon nanostructure of a graphene / silicon biosensor according to an embodiment of the present invention.

Referring to FIG. 3, the surface (Si NW surface) of the silicon nanostructure 120 of the graphene / silicon biosensor 100 according to an embodiment of the present invention is formed of an element and a negatively charged hydroxide ion (OH-) (Hydroxide). &Lt; / RTI &gt;

In addition, the surface of the silicon nanostructure 120 may be coated with a functional material that improves the adhesion so that DNA adheres well, and the functional material may include 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde (Glutaraldehyde).

(3-APTES) material containing NH 2 may be coated on the surface of the silicon nanostructure 120, and the 3-aminopropyltriethoxysilane (3-APTES) material containing NH 3 may be coated on the surface of the silicon nanostructure 120. A glutaraldehyde material having a molecular formula of C 5 H 8 O 2 may be coated on the surface of the silicon nanostructure 120 coated with the APTES material.

For example, the 3-aminopropyltriethoxysilane (3-APTES) material produces a strong covalent bond between the surfaces of the silicon nanostructure 120 and a 3-aminopropyltriethoxysilane (3-APTES) The surface of the coated silicon nanostructure 120 can be formed with a structure in which the surface of the silicon nanostructure 120 is firmly fixed by heat treatment for hydrolysis.

Also, the glutaraldehyde substance is a chemically modifying agent of a protein that reacts with and bridge with an amino group (-NH 2 ), and is coated on the surface of the silicon nanostructure 120 to form a 3-aminopropyltriethoxysilane (3-APTES) substance, it is possible to increase the binding force with the probe DNA.

3, the surface of each of the plurality of silicon nanostructures 120 constituting the graphene / silicon biosensor 100 is coated with 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde (Glutaraldehyde), it is possible to improve the adhesion of DNA to be discriminated.

Hereinafter, the process of identifying DNA using the graphene / silicon biosensor will be described in detail with reference to FIGS. 4A to 4C.

4A to 4C show examples of the mechanism of a graphene / silicon biosensor according to an embodiment of the present invention.

Referring to FIG. 4A, a probe DNA, a target DNA, and a dummy DNA can be used to detect DNA using a DNA detection apparatus using a graphene / silicon biosensor.

According to an embodiment, the probe DNA is a single stranded DNA including a nucleotide sequence of a DENND2D promoter, which is a human gene, for example, 5'-tta-gcg-cgg-agt-tgg -gag-cgg-gag-tcg &lt; / RTI &gt;

In addition, the target DNA is a single stranded DNA that forms a pair of the probe DNA and the nucleotide sequence. For example, the nucleotide sequence of aat-cgc-ccg-act-cca-ctc-ccg-ctc-cga- Lt; / RTI &gt; DNA.

In addition, the dummy DNA is a single stranded DNA which forms a nucleotide sequence pair completely different from the probe DNA. For example, the base sequence of 5'-tct-tgc-aca-agt-tta-aga-ggg-aaa-gga Lt; / RTI &gt; DNA.

Referring to FIG. 4B, a probe DNA extracted from a DENND2D promoter, which is a human gene, is applied to a graphene / silicon biosensor and can be attached to the surface of a plurality of silicon nanostructures 120.

According to the embodiment, on the surface of the silicon nanostructure 120, sample DNA extracted from various sample cells in addition to the above-mentioned probe DNA can be attached. In addition, the probe DNA (probe DNA) may refer to all of the single-stranded DNAs of the sample DNA to be detected in addition to the probe DNA, and the kind thereof is not limited.

Referring to FIG. 4C, at least one of the target DNA and the dummy DNA may be applied to a graphene / silicon biosensor to which the probe DNA extracted from the DENND2D promoter is attached.

A DNA detection apparatus using a graphene / silicon biosensor can be applied to a graphene / silicon biosensor in which probe DNA is attached to the surface of the silicon nanostructure 120 by applying at least one of target DNA and dummy DNA to be measured, It is possible to detect the adhesion between the DNA and the target DNA and the dummy DNA.

For example, a probe DNA that is a single stranded DNA having a nucleotide sequence in the sample DNA extracted from the DENND2D promoter is applied and attached to a graphene / silicon biosensor, and a measurement target (for example, a patient or a clinical subject ) Can be applied to a graphene / silicon biosensor having a probe DNA attached thereto, in which single-stranded DNA having a nucleotide sequence in the target DNA extracted from the target cells of the target DNA.

Here, when the DNA of the single strand having the nucleotide sequence of the target DNA is a target DNA that forms a pair of the probe DNA and the nucleotide sequence extracted from the DENND2D promoter, the probe DNA and the target DNA may be attached to each other, but the nucleotide sequence If the DNA of a single helix is a dummy DNA that forms a completely different nucleotide sequence from the probe DNA, the probe DNA and the dummy DNA may not be attached.

Therefore, the DNA detection apparatus using the graphene / silicon biosensor according to the embodiment of the present invention can identify the DNA depending on whether the probe DNA is attached to the target DNA or the dummy DNA.

5 is a block diagram of a DNA detection apparatus using a graphene / silicon biosensor according to an embodiment of the present invention.

Referring to FIG. 5, a DNA detection apparatus 200 using a graphene / silicon biosensor according to an embodiment of the present invention includes a graphene / silicon biosensor 100 and a detection unit 210.

The graphene / silicon biosensor 100 comprises a substrate, a functional material comprising at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde bonded in alignment with the substrate, A plurality of silicon nanostructures formed on the silicon nanostructure, a graphene layer disposed on the silicon nanostructure, and an electrode formed on the bottom of the substrate and on the top of the graphene layer, respectively.

The structure and structure of the graphene / silicon biosensor 100 are described in FIG. 1 and will not be described here.

The detection unit 210 identifies the DNA according to whether a target DNA pair forming a pair with the predetermined probe DNA attached to the surface of the silicon nanostructure through the functional material is attached.

For example, the detection unit 210 of the DNA detection apparatus using the graphene / silicon biosensor according to an embodiment of the present invention may include a graphene / silicon biosensor having a single-stranded probe DNA-containing nucleotide sequence extracted from the DENND2D promoter, A single-stranded target DNA paired with the probe DNA may be applied to the silicon biosensor 100 to detect whether the target DNA is attached to the probe DNA.

The probe DNA may be a single stranded DNA comprising a nucleotide sequence including a nucleotide sequence of a sample DNA extracted from a sample cell, and the target DNA may be a single stranded DNA strand that is paired with a nucleotide sequence Can be referred to as DNA.

That is, the target DNA may be a single strand of DNA extracted from a cell to be measured.

The detection unit 210 can detect the DNA by detecting a change in the amount of current depending on whether the probe DNA and the target DNA are attached from the electrode included in the graphene / silicon biosensor 100.

For example, the detection unit 210 can identify whether adhesion between the probe DNA and the target DNA is based on an increase in the amount of current when the target DNA paired with the probe DNA is applied. If a dummy DNA that is not paired with the probe DNA is applied to the graphene / silicon biosensor 100 to which the probe DNA is attached, the amount of current is rapidly decreased and the degree of reactivity is very low. ) Can identify whether DNA is attached or not.

FIGS. 6A to 6D show scanning electron microscopy (SEM) images of a graphene / silicon biosensor according to an embodiment of the present invention.

6A to 6D are cross-sectional views of a graphene / silicon biosensor according to an embodiment of the present invention shown in FIGS. 2A to 2D, FIG.

6A is a scanning electron microscope image of a gold mesh formed on a substrate formed of silicon as shown in FIG. 2A. Referring to FIG. 6A, a porous gold mesh array is formed of silicon (Si). &Lt; / RTI &gt;

FIG. 6B is a scanning electron microscope image of a plurality of silicon nanostructures formed on a substrate as shown in FIG. 2B. Referring to FIG. 6B, it can be seen that a plurality of silicon nanostructures are formed.

FIGS. 6C and 6D are SEM images of a front surface and a side surface of a graphen / silicon biosensor including a substrate, a silicon nanostructure formed on the substrate, and a graphene layer formed on the silicon nanostructure, 6C and 6D, a plurality of silicon nano structures (Si NWs) having a constant height are arranged on a substrate formed of silicon (Si), and a graphene layer covering a partial area of a plurality of silicon nano structures .

FIG. 7A is a graph showing the current change curve with respect to the concentration of the probe DNA according to the embodiment of the present invention, and FIG. 7B is a graph showing the current change according to the attachment of the target DNA and the dummy DNA to the probe DNA according to the embodiment of the present invention Curve and response diagrams.

More specifically, FIG. 7A shows the current change according to the concentration of single-stranded probe DNA containing a nucleotide sequence of DNA extracted from the DENND2D promoter.

Referring to FIG. 7A, it can be seen that the amount of current flowing through the graphene / silicon biosensor increases as the concentration of the probe DNA increases. When the concentration of the probe DNA is about 500 nM, the amount of current indicates saturation .

FIG. 7B is a graph showing the relationship between target DNA and dummy DNA in a graphene / silicon biosensor having a probe DNA attached to a single helix including a nucleotide sequence of DNA extracted from a DENND2D promoter, In the case of applying the coating solution.

Referring to FIG. 7B, when the target DNA paired with the probe DNA is applied to the graphene / silicon biosensor coated with the probe DNA, the amount of current flowing through the graphene / silicon biosensor It can be confirmed that it increases sharply.

However, when the dummy DNA not attached to the probe DNA is applied to the graphene / silicon biosensor coated with the probe DNA, it can be confirmed that the amount of current flowing through the graphene / silicon biosensor is reduced.

In addition, as shown in FIG. 7B, it can be seen that the degree of reactivity when the target DNA paired with the probe DNA is applied varies by about 0.6 mA in absolute value, and it is found that the degree of reactivity changes to about 120%.

Therefore, the graphene / silicon biosensor according to the embodiment of the present invention can identify the DNA by detecting the amount of current depending on whether the probe DNA, the target DNA, or the dummy DNA is attached.

FIG. 8 is a graph showing a result of a recycling characteristic of a graphene / silicon biosensor according to an embodiment of the present invention.

More specifically, FIG. 8 is a graph showing the change in the characteristics of a graphene / silicon biosensor according to an embodiment of the present invention, The DNA attached to the surface is removed, and the DNA detection characteristics of the graphene / silicon biosensor are evaluated again.

Referring to FIG. 8, the graphene / silicon biosensor according to an embodiment of the present invention performs a stable DNA detection function at about 0.1% of the target DNA at a minimum concentration of about 20% while driving a recycling cycle of 10 times . Therefore, it can be seen that the graphene / silicon biosensor according to the embodiment of the present invention is easy to be recycled.

FIGS. 9A to 9C illustrate fluorescence characteristic images according to the adhesion of DNA applied to a graphene / silicon biosensor according to an embodiment of the present invention.

More specifically, FIGS. 9A to 9C illustrate a method of applying a probe DNA, a target DNA, and a dummy DNA having fluorescence colors to a graphene / silicon biosensor according to an embodiment of the present invention, It is an image observed with a microscope.

As shown in FIG. 9A, the reddish probe DNA was applied to a graphene / silicon biosensor, and the surface of the silicon nanostructure was observed under a fluorescence microscope. As a result, it was found that the probe DNA adhered well to the surface of the silicon nanostructure Can be confirmed.

As shown in FIG. 9B, the target DNA having an orange color was applied to a graphene / silicon biosensor having a probe DNA attached thereto, and the surface of the silicon nanostructure was observed under a fluorescence microscope. As a result, It can be confirmed that the silicon nanostructure adheres well to the surface of the attached silicon nanostructure.

9C, dummy DNA having a green color was applied to a graphene / silicon biosensor having a probe DNA attached thereto, and the surface of the silicon nanostructure was observed with a fluorescence microscope. As a result, It can be confirmed that they do not adhere well to the surface of the silicon nanostructure and are deposited and precipitated with each other.

9a to 9c, the target DNA pairing with the nucleotide sequence of the probe DNA shows a high affinity with the probe DNA, whereas the dummy DNA having completely different nucleotide sequence pairs is not attached to the probe DNA Thus, more accurate DNA can be identified from the graphene / silicon biosensor using the adhesion characteristics between these DNAs.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. For example, it is to be understood that the techniques described may be performed in a different order than the described methods, and / or that components of the described systems, structures, devices, circuits, Lt; / RTI &gt; or equivalents, even if it is replaced or replaced.

Therefore, other implementations, other embodiments, and equivalents to the claims are also within the scope of the following claims.

100: Graphene / Silicon Biosensor
110: substrate
120: Silicon nanostructure
130: graphene layer
140a, 140b:

Claims (6)

Board;
A plurality of silicon nanostructures aligned and bonded to the substrate and coated with a functional material including at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde;
A graphene layer disposed on the silicon nanostructure; And
Electrodes formed on the lower portion of the substrate and the upper portion of the graphene layer,
/ RTI &gt;
Detecting a change in an amount of current depending on whether or not a target DNA (pair of target DNAs) is attached to a predetermined probe DNA attached to the surface of the silicon nanostructure through the functional material,
The surface of the silicon nanostructure is
Generating a covalent bond and to the 3-aminopropyl containing silane and NH 2, an amino group (-NH 2) and a chemical modification reaction agent of the glutaraldehyde binding proteins bridge is coupled to the coupling force between the probe DNA Graphene / Silicon biosensor to increase. The method according to claim 1,
The silicon nanostructure includes
Wherein the functional material for binding the probe DNA is coated on the graphene / silicon biosensor. The method according to claim 1,
The functional material
Wherein the graphene / silicon biosensor is a material that attaches the probe DNA to the surface of the silicon nanostructure by generating a strong covalent bond between the surfaces of the silicon nanostructure. The method according to claim 1,
The electrode
Wherein the graphene / silicon biosensor is formed of at least one material selected from silver (Ag), gold (Au), copper (Cu), aluminum (Al), platinum (Pt) and alloys thereof. A plurality of silicon nanostructures which are aligned and bonded to the substrate and coated with a functional material including at least one of 3-aminopropyltriethoxysilane (3-APTES) and glutaraldehyde; A graphene layer disposed on the upper portion of the structure, and an electrode formed on the lower portion of the substrate and on the upper portion of the graphene layer, respectively; And
And a detector for detecting a change in an amount of current depending on the presence or absence of a pair of target DNAs attached to a predetermined probe DNA attached to the surface of the silicon nanostructure through the functional material to identify the DNA,
The surface of the silicon nanostructure is
Generating a covalent bond and to the 3-aminopropyl containing silane and NH 2, an amino group (-NH 2) and a chemical modification reaction agent of the glutaraldehyde binding proteins bridge is coupled to the coupling force between the probe DNA To heighten
DNA detection device using graphene / silicon biosensor. delete
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11415577B2 (en) 2018-10-31 2022-08-16 Electronics And Telecommunications Research Institute Method of detecting bio-material
WO2024076193A1 (en) * 2022-10-06 2024-04-11 한국생명공학연구원 Localized surface plasmon resonance sensor with improved sensitivity and manufacturing method therefor
WO2024080388A1 (en) * 2022-10-11 2024-04-18 엘지전자 주식회사 Biosensor

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Jing Tang et al., "Sensitive enzymatic glucose detection by TiO₂nanowire photoelectrochemical biosensors", JOURNAL OF MATERIALS CHEMISTRY A, 2, pp.6153-6157, 2014*
Jungkil Kim et al., "Graphene/Si-nanowire heterostructure molecular sensor", SCIENTIFIC REPORTS 4, pp.1-4, 2014*

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11415577B2 (en) 2018-10-31 2022-08-16 Electronics And Telecommunications Research Institute Method of detecting bio-material
WO2024076193A1 (en) * 2022-10-06 2024-04-11 한국생명공학연구원 Localized surface plasmon resonance sensor with improved sensitivity and manufacturing method therefor
KR20240048301A (en) * 2022-10-06 2024-04-15 한국생명공학연구원 Localized surface plasmon resonance sensor with improved sensitivity and manufacturing method thereof
KR102665267B1 (en) 2022-10-06 2024-05-13 한국생명공학연구원 Localized surface plasmon resonance sensor with improved sensitivity and manufacturing method thereof
WO2024080388A1 (en) * 2022-10-11 2024-04-18 엘지전자 주식회사 Biosensor

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