WO2015025993A1

WO2015025993A1 - System and method for analyzing molecular sequence in real time using stacked nano-ribbon

Info

Publication number: WO2015025993A1
Application number: PCT/KR2013/007477
Authority: WO
Inventors: 최중범
Original assignee: 나노칩스(주)
Priority date: 2013-08-21
Filing date: 2013-08-21
Publication date: 2015-02-26

Abstract

The present invention relates to a system for analyzing a molecular sequence which enables the analysis in real time of a unit molecular sequence forming a bio-polymer by using a stacked nano-ribbon, and is comprised of nanopores, a nano-ribbon, measuring electrodes, an insulation layer, and a substrate. The measuring electrodes are electrically connected to the nano-ribbon at both ends in a lengthwise direction of the nano-ribbon, a current made to flow in the nano-ribbon by means of a predetermined voltage applied through the measuring electrodes is changed by the respectively different inherent electric dipoles of unit molecules of bio-polymer which passes nanopores formed in the middle of the nano-ribbon, and the degrees of the changes are detected, so that the sequences of the unit molecules forming the bio-polymer can be analyzed in real time.

Description

Real-Time Molecular Sequence Analysis System and Analysis Method Using Stacked Nanoribbons

The present invention relates to a real-time molecular sequence analysis system using stacked nanoribbons, and more particularly, at least four nanoribbons are stacked on a substrate with an insulating layer interposed therebetween, the substrate, the insulating layer and the nano A nanopore is formed penetrating the center of the ribbon in a vertical direction, and the nanoribbons are formed by the interaction between the unit molecules and the nanoribbons constituting the biopolymer while the biopolymer passes through the nanopore. A molecular sequence analysis system that detects the change of the unit spherical component in real-time by sensing the current flowing through the measured and measured change through a measurement electrode formed at each end of the longitudinal length of the nanoribbon, and It's about how.

Decoding the unit molecule sequence (amino acid molecule sequence of the protein, DNA base molecule sequence) constituting the biological polymer means decoding the biometric information, the molecular genetic understanding can make early diagnosis of human disease It is very important in that it is.

For example, DNA is composed of nucleotide units, and each nucleotide has the same one deoxyribose and phosphate group and has four different bases: adenine (A), guanine (G), and cytosine (C). ), Thymine (T) is present, and A and G are Purine series having two cyclic structures, and C and T are Pyrimidine series having one cyclic structure.

Based on the nucleotides recorded in the deoxyribonucleic acid, messenger RNA (mRNA) is transcribed and proteins are synthesized through the binding of amino-acids in ribosomes. If there is a mutated nucleotide sequence different from the original nucleotide sequence, protein synthesis may not be possible or abnormal proteins may be synthesized and expressed, resulting in physiological problems. For this reason, the meaning of deciphering the information possessed by DNA is very important from a pathological point of view for the early diagnosis and treatment of diseases.

Various methods have been developed for DNA sequencing, ranging from early Maxam-Gilbert sequencing to Dye-Terminator sequencing. However, these methods limit the length of DNA that can be analyzed per unit time, and require a long preparation process such as repeating DNA amplification, radioisotope substitution and staining for accurate analysis.

In addition, the analysis of the entire DNA through the pieces of DNA is done through the processing of a lot of data, and requires a lot of money because of the time required and the chemicals required for analysis. The problem faced by this analytical method is that various solutions have been proposed through the convergence of rapidly developed nanotechnology and biotechnology, and can provide infinite potential technology.

As a representative example, electrical analysis methods have been proposed by applying a solid element that can be used semi-permanently by breaking away from conventional chemical reaction methods. Successful implementation of these technologies can reduce the time required for preparation and analysis, reduce waste by reducing chemicals, and enable simpler and more accurate analysis.

Therefore, the present invention has been made to solve the conventional problems as described above, according to an embodiment of the present invention, to solve the problems of the conventional unit molecular sequence analysis system using nanotechnology, molecular sequence analysis It reduces excessive time and harmful chemicals that are invested in preliminary work, and enables real-time unit molecular analysis at high speed using semi-permanent solid elements, unlike conventional methods, which provides a cheaper molecular sequence analysis system. For the purpose of

In addition, according to one embodiment of the present invention, including the movement means having a piezoelectric element inherent electricity of different unit molecules constituting the biopolymer while controlling the movement speed of the biopolymer (biological polymer) passing through the nanopore A real-time molecular sequence analysis system and method using stacked nanoribbons with piezoelectric elements that sense the change in charge distribution and current induced from dipoles or intrinsic energy orbits and decode the identity of unit spherical components in real time through nanochannels. It aims to provide.

Other objects, specific advantages and novel features of the present invention will become more apparent from the following detailed description and preferred embodiments in conjunction with the accompanying drawings.

An object of the present invention is a nanoribbon having a specific width in which a current change is induced by interaction with the nearest unit molecules of the biomolecules; Measuring electrodes formed at each of both ends of the longitudinal direction of the nanoribbon to measure a change in current; An insulating layer formed on the upper and lower surfaces of the nanoribbons to electrically insulate the nanoribbons from each other; And nanopores that penetrate the insulating layer and the nanoribbons in the vertical direction and allow unit molecules forming the biopolymer to pass through. The real-time molecular sequence analysis system using the stacked nanoribbons may include the nanopores.

The apparatus may further include a substrate for supporting the nanoribbons, the measuring electrode, and the insulating layer, and the nanopores may be formed by penetrating the substrate, the insulating layer, and each of the nanoribbons in a vertical direction.

Four or more nanoribbons may be configured to be stacked on a substrate with an insulating layer interposed therebetween.

Each nanoribbon may have a unit molecule coating part coated with different unit molecules at a position where the nanopores are formed to induce complementary bonding between the unit molecule and the coating part passing through the nanopores.

Nanoribbons include zig-zag graphene or two-dimensional topological insulators, so the inside of the nanoribbon is characterized by being electrically insulating and having edges that are conductive. Can be.

The measuring electrode may be made of a conductor including at least one of gold, silver, copper, platinum, palladium, titanium, nickel, and cobalt, and may be electrically connected to the nanoribbon.

The insulating layer consists of an electrical insulator comprising at least one of a silicon oxide film, a silicon nitride film, an aluminum oxide film, a silver oxide film, a zinc oxide film, and a hafnium oxide film, and insulates the nanoribbons from the electrolyte and other nanoribbons. It can be characterized.

At the edge of the nanoribbon in which the nanopores are formed, a hydrogen atom or a nitrogen atom may be bonded to a carbon atom having a dangling bond.

The thickness of the nanoribbons may be characterized in that less than 5 kPa.

The width of the nanoribbons may be characterized in that less than 10nm.

The diameter of the nanopores may be characterized in that less than 10nm.

The method may further include a voltage applying unit for applying a voltage to the measurement electrode.

The method may further include analyzing means for acquiring the current change data based on the current measured by the measuring electrode, and determining the sequence of the unit molecules of the biopolymer based on the current change data.

In another category, an object of the present invention is to apply a voltage to a measurement electrode connected to both ends of a length of each nanoribbon such that a current flows through the nanoribbon; Applying a voltage difference or a fluid pressure difference between one side and the other side of the substrate to allow the biopolymer to pass through the nanopores; Changing the current flowing through the nanoribbons by interacting with the nanoribbons of the unit molecules forming the biopolymer; Detecting a change in current flowing through the nanoribbon; And determining the sequence of unit molecules by combining and analyzing the current change flowing through each nanoribbon. This may be achieved as a real-time molecular sequence analysis method using stacked nanoribbons.

The current flowing through the nanoribbon may be characterized as including an edge current.

In the step of changing the current, the nanoribbons are stacked on the substrate with four or more insulating layers interposed therebetween, and each nanoribbon is provided with unit molecular coatings coated with different unit molecules at positions where nanopores are formed. , Complementary bonding of the unit molecule and the coating portion passing through the nanopores may be characterized in that the current flowing through the nanoribbons is changed.

The detecting and determining may be based on a change in current flowing through the nanoribbons detected by the measuring electrode, and the analysis means may identify the sequence of unit molecules by combining and analyzing the change in current flowing through the nanoribbons. Can be.

According to an embodiment of the present invention, nanoribbons are stacked on a substrate with an insulating layer interposed therebetween, and each unit molecule of a biopolymer that moves nanopores vertically penetrating the center of the substrate, the insulating layer, and the nanoribbons By detecting the change in current of the nanoribbon from the dipole in real time through the measuring electrode electrically connected to the nanoribbon, it analyzes the identity of each unit molecule. The use of these molecules can be greatly reduced, and the unit molecular sequence can be analyzed at high speed and precision.

In particular, in order to analyze the sequence of ss-DNA, at least four layers of nanoribbons are sandwiched between insulating layers, and four different DNA base molecules (A and G) are present in each nanoribbon of nanopores formed at the center thereof. , T, C) can be maximized the detection efficiency and reliability by maximizing the current change of the nanoribbon through complementary bonding with base molecules moving into the nanopore, and through a single ss-DNA movement 4 All nucleotide sequences consisting of two base molecules can be analyzed.

In addition, according to an embodiment of the present invention, including a moving means having a piezoelectric element, by attaching ss-DNA which is an example of a biopolymer to the probe to pass through the nanochannel by driving the moving unit and the tension, compression of the piezoelectric element. By controlling the moving speed of the ss-DNA has an effect that can more accurately decode the ss-DNA base sequence.

Although the present invention has been described in connection with the above-mentioned preferred embodiments, it will be readily apparent to those skilled in the art that various other modifications and variations are possible without departing from the spirit and scope of the present invention. Are all within the scope of the appended claims.

1 is a perspective view of a real-time molecular sequence analysis system using stacked nanoribbons applied to DNA base molecule sequence decoding according to an embodiment of the present invention,

2 is a cross-sectional view taken along line A-A of FIG.

3 is a partial cross-sectional view of a real-time molecular sequence analysis system using a laminated nanoribbon having a piezoelectric element according to an embodiment of the present invention;

4 is a flowchart of a method for real-time molecular sequence analysis using stacked nanoribbons according to an embodiment of the present invention;

FIG. 5 is a graph showing current change data measured using nanoribbons coated with four different bases applicable according to one embodiment of the present invention; FIG.

6 is a flowchart illustrating a real-time molecular sequence analysis method using stacked nanoribbons having piezoelectric elements according to an embodiment of the present invention.

4: measuring tank 5: electrode element

10: ss-DNA 40: Head

50: moving part 50: moving part

60: piezoelectric element 70: probe

100: nanopores

200: nanoribbon 200A: A-coated nanoribbon

200G: G-coated nanoribbons 200T: T-coated nanoribbons

200C: C coated nanoribbons 300: measuring electrode

400: insulating layer 500: substrate

DETAILED DESCRIPTION Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, in describing in detail the operating principle of the preferred embodiment of the present invention, if it is determined that the detailed description of the related known functions or configurations may unnecessarily obscure the subject matter of the present invention, the detailed description thereof will be omitted.

In addition, the same reference numerals are used for parts having similar functions and functions throughout the drawings. Throughout the specification, when a part is connected to another part, this includes not only the case where it is directly connected, but also the case where it is indirectly connected with another element in between. In addition, the inclusion of any component does not exclude other components unless specifically stated otherwise, it means that may further include other components.

In describing the embodiments of the present invention, descriptions of well-known configurations that will be obvious to those skilled in the art will be omitted so as not to obscure the subject matter of the present invention. In addition, when referring to the drawings, it should be considered that the thickness of the lines shown in the drawings, the size of the components, etc. may be exaggerated for the sake of clarity and convenience of the description. The term is based on the direction shown in the drawings unless otherwise noted.

The real-time molecular sequence analysis system using the stacked nanoribbons according to the present invention decodes the unit molecule sequence (for example, amino acid molecules of proteins, or base molecule sequences of DNA, etc.) constituting various biopolymers such as proteins or DNA. It can be used to As a specific example, specific contents applied to DNA nucleotide sequence analysis will be described below.

Hereinafter will be described the configuration and function of a real-time molecular sequence analysis system using a laminated nanoribbon according to an embodiment of the present invention. First, FIG. 1 illustrates a perspective view of a real-time molecular sequence analysis system using stacked nanoribbons applied to DNA base molecule sequence decoding according to an embodiment of the present invention, and FIG. 2 is a cross-sectional view of FIG.

As shown in Figure 1 and 2, the real-time molecular sequence analysis system using a stacked nanoribbon according to an embodiment of the present invention is largely nanopore 100, nanoribbon (200: 200A, 200G, 200T, 200C) , The measurement electrode 300, the insulating layer 400, the substrate 500, and the like, may be included.

Referring to the basic function of the present invention having the above configuration, a predetermined voltage is applied to the measurement electrode 300 to flow a predetermined current through the nanoribbons 200, and the other side of the substrate 500 Single-stranded DNA (hereinafter referred to as ss-DNA) by a voltage difference or a fluid pressure difference applied between (between the upper side and the lower side of the substrate 500 as shown in FIGS. 1 and 2), 10 passes through the nanopores (100).

From the different nucleotide-specific electric dipoles that make up the ss-DNA 10 passing through the nanopores 100, the charge distribution of the nanoribbons 200 around the nanopores 100 is induced and from these changes in charge distribution, The sequence of nucleotides constituting the ss-DNA 10 may be analyzed by analyzing a change in edge current flowing through the edge of the ribbon 200.

In addition, as shown in Figure 2, at the edge of each of the nanoribbon 200 at the position where the nanopore 100 is formed is coated with any one of the different bases A, G, T, C through the nanopore 100 By maximizing the interaction with the nanoribbons 200 through the complementary bond (AT or GC) nucleotides ss-DNA (10) to maximize the current change flowing through the edge of the nanoribbons (200).

That is, as shown in Figure 2, in the real-time molecular sequence analysis system using a laminated nanoribbon according to an embodiment of the present invention, four nanoribbons 200, the insulating layer 400 between the substrate 500 It can be seen that it is stacked on the configuration, based on the shown in Figure 2 based on the nanoribbon (200) located at the top of the nanopore 100 is located on both ends of the edge of the nanoribbon coated nanoribbon (200A) This is provided, the second from the top is G nano-coated nanoribbons (200G) is provided, the third from the top T-coated nanoribbons (200T), the fourth from the top is the C-base coated nanoribbons ( It can be seen that 200C) is provided.

The number of structures and nanoribbons coated with these different bases is not limited to the exemplary embodiments of the present invention and drawings, but may be composed of more nanoribbons, within the range including the technical spirit of the present invention. As long as a person skilled in the art can easily substitute and change, it should be interpreted within the scope of the present invention.

In addition, the configuration of the real-time molecular sequence analysis system using the stacked nanoribbons according to an embodiment of the present invention as described above in detail for each component as follows.

First, as shown in FIGS. 1 and 2, the nanopores 100 are formed to vertically penetrate through the centers of the substrate 500, the plurality of insulating layers 400, and the plurality of nanoribbons 200. , ss-DNA (10) has a diameter that can pass without twisting or overlapping, usually in the range of 2nm or less. In addition, the ss-DNA 10 passes the nanopores 100 by a voltage difference or a fluid pressure difference applied between one side and the other side of the substrate 500.

Here, ss-DNA (10) is used to induce a change in the charge distribution of the nanoribbons 200 from the electric dipoles of different nucleotides by exposing the base to the outside, DNA is a double helix structure for one strand Since the other strand has a complementary sequence, it is possible to grasp the entire DNA structure by analyzing the nucleotide sequence with one ss-DNA (10).

The nanoribbons 200 are used to detect the actual nucleotides. Since the distance between the different bases forming the ss-DNA 10 is 5 m or less, the thickness of the nanoribbons 200 should also be 5 m or less. When analyzing nucleotides it is possible not to be greatly interfered with other adjacent nucleotides.

In addition, in order to induce a change in the effective current of the nanoribbons 200 from the base electric dipole should have a width of less than 10nm. In addition, zig-zag graphene or two-dimensional topological insulators satisfying the above conditions may serve as the nanoribbons 200.

In the nanoribbons shown in FIGS. 1 and 2, 200A is coated with deoxyribonucleotide (dATP) having adenine or adenine as a base on the

nanopore

100, and 200G is similarly deoxyribonucleotide having a guanine or guanine as a base. dGTP) is coated with thymine or deoxyribonucleotide (dTTP) with thymine as base, and 200C is coated with deoxyribonucleotide (dCTP) with cytosine or cytosine as base.

Here, when the ss-DNA 10 passes through the nanopores 100, the nanoribbons through the complementary bond (AT or CG) of the base constituting the ss-DNA 10 and the base coated on the nanoribbons 200. The change of the charge distribution is maximized, and thus the change of the edge current of the nanoribbon is maximized. In addition, by combining hydrogen atoms or nitrogen atoms with carbon atoms having “dangling bonds” at the edges of the nanopores 100 and nanoribbons 200 in addition to the coating of nucleotides, constituent unit molecules and nanoribbons of biopolymers You can also maximize your interaction.

In addition, the measuring electrode 300 according to an embodiment of the present invention may be composed of a conductor including gold, silver, copper, platinum, palladium, titanium, nickel, cobalt, the longitudinal direction of the nanoribbon 200 It is electrically connected to both ends, and the voltage is applied by the voltage applying unit.

In addition, the stacked nanoribbons 200 are formed on the upper and lower surfaces of the nanoribbons 200 for the analysis of each nucleotide, and an insulating layer 400, which is an electrical non-conductor, is formed in a length direction with one nanoribbon 200. The measuring electrodes 300 formed at both ends are electrically insulated from an external electrolyte or other nanoribbons 200. In addition, the substrate 500 is mechanically supported by the nanoribbons 200 on which the insulating layer 400 and the measurement electrode 300 are formed.

In addition, the real-time molecular sequence analysis system using a laminated nanoribbon according to an embodiment of the present invention includes an electrode element 5, a probe 70, a piezoelectric element 60, a moving unit 50, a control means, The movement speed of the ss-DNA 10 passing through the nanopores can be controlled.

Figure 3 shows a cross-sectional view of a real-time molecular sequence analysis system using a stacked nanoribbons having a piezoelectric element according to an embodiment of the present invention. As shown in FIG. 3, it can be seen that the substrate 500 serves as a separation wall separating both ends of the measuring tank 4 in which the electrolyte for maintaining the ss-DNA 10 is maintained therein.

The electrode elements 5 are installed at both sides of the substrate at specific intervals to form a potential difference at both ends of the substrate 500 so that the ss-DNA 10 can be supplied to move the nanopores 100. do. That is, the electrode elements 5 are provided in pairs and installed at the upper end and the lower end of the measuring bath 4, respectively. Then, the voltage is supplied to the electrode element 5 so that the lower end has a + pole and the upper end has a-pole, thereby forming a potential difference between the upper end and the lower end of the substrate 500. Therefore, since the ss-DNA 10 is basically -charged, the ss-DNA 10 can be moved from the upper end to the lower end through the nanopores 100 by this potential difference.

The real-time molecular sequence analysis system using the stacked nanoribbons having the piezoelectric element 60 according to the embodiment of the present invention further includes a moving means. The moving means includes a head 40 installed on one side of the measuring tank 4 and a moving part 50 moving in the x, y, and z axis directions with respect to the head, and a piezoelectric element 60 that is tensioned or compressed by an applied voltage. And, it may be provided as a control means (not shown) for controlling the driving of the piezoelectric element 60 and the moving unit 50.

The moving unit 50 moves the piezoelectric element 60 and the probe 70 in the x-axis, the y-axis, and the z-axis in the vertical direction with respect to the head 40 at a level of 0.1 to several tens of micrometers. The moving unit 50 may be composed of a piezo motor or the like. In addition, the piezoelectric element 60 is tensioned or compressed in the z-axis, x-axis, and y-axis directions in the range of 0.1 to 10 μm by the applied voltage.

Accordingly, the ss-DNA 10 is nanopored by driving the piezoelectric element 60 and the moving unit 50 by bonding the end of the ss-DNA 10 to the end surface of the probe 70. It is possible to adjust the moving speed passing through (100).

Hereinafter, a brief description of a real-time molecular sequence analysis method using the stacked nanoribbons 200 according to an embodiment of the present invention. This analysis method uses a real-time molecular sequence analysis system using the stacked nanoribbons mentioned above. 4 is a flowchart illustrating a real-time molecular sequence analysis method using stacked nanoribbons according to an embodiment of the present invention.

First, a voltage is applied to the measurement electrodes 300 provided at both ends of each nanoribbon 200 by a voltage applying unit so that a predetermined current flows through the nanoribbon 200 (S10). Then, a voltage difference or a fluid pressure difference is applied between one surface and the other surface of the substrate 500 to allow the biopolymer to pass through the nanopores 100 (S20).

In addition, the unit molecules forming the biopolymer interact with each nanoribbon 200 to change a current flowing through the nanoribbon 200. At this time, most of the current flowing through the nanoribbons 200 corresponds to an edge current. In addition, the change of the current flowing through the nanoribbons 200 is sensed by the measuring electrodes 300 provided at both ends of each of the nanoribbons 200 (S30). In this case, the nanoribbons 200 are stacked on the substrate 500 with four or more insulating layers 400 interposed therebetween, and each of the nanoribbons 200 has different units at positions where the nanopores 100 are formed. Comprising a unit molecule coating unit coated with a molecule, the complementary bonding of the unit molecule and the base coating portion passing through the nanopores 100 can be induced to maximize the current change flowing through the nanoribbons 200.

In addition, the analysis means obtains the current change data by the unit molecules constituting the biopolymer passing through the nano-pores (100) (S40). In addition, the analyzing means may determine the sequence of the unit molecules by combining and analyzing the obtained current change data (S50).

That is, based on the current change flowing through the nanoribbons 200 sensed by the measuring electrode 300, the analysis means identifies the sequence of unit molecules by combining and analyzing the current change data flowing through the nanoribbons 200. Done.

FIG. 5 is a graph showing current change data measured in real-time using nanoribbons 200 coated with four different bases applicable according to an embodiment of the present invention. That is, FIG. 5 shows four independent nanoribbons 200 coated with ss-DNA 10 having the same base sequence CTGACTGA ... as in FIG. 2 passing through the nanopore 100 with each other base. Shows the measured data in real time that can be predicted by edge current modulation.

The graph shown at the top left in FIG. 5 is the current change data measured in the nanoribbons 200A coated with base A, and the graph shown at the top right is the current change measured in the nanoribbons 200G coated with base G. Data, the graph shown at the bottom left is a base T coated nanoribbons 200T, and the graph shown at the bottom right is a base C coated nanoribbons 200C.

Therefore, the analytical means can analyze the base sequence of the ss-DNA 10 that passed through the nanochannel by combining-analyzing these four data in real time, and if the analysis of the graph shown in FIG. It can be read that the sequence becomes CTGACTGA ... in real time as in FIG. 2.

In addition, as mentioned above, the real-time molecular sequence analysis system using the stacked nanoribbons according to an embodiment of the present invention, the electrode element 5, the probe 70, the piezoelectric element 60, the moving part 50, Including the control means, it is possible to adjust the moving speed of the ss-DNA 10 passing through the nanopores.

Hereinafter, a real-time molecular sequence analysis method using the stacked nanoribbons having such piezoelectric elements will be described. First, FIG. 6 illustrates a flowchart of a real-time molecular sequence analysis method using stacked nanoribbons having piezoelectric elements.

First, the control means drives the moving part 50 of the moving means to attach the end of the ss-DNA 10 to the end surface of the probe 70 while moving the moving part 50 in the x, y and z axis directions. It becomes (S100).

Then, the control means drives the moving unit 50 to adjust the direction and position the end of the probe 70 close to the nano-pores (100) (S200). In addition, the step of attaching the end of the ss-DNA (10) to the end surface of the probe 70 (S100) and the position of the ss-DNA (10) in close proximity to the nanopore 100 (S200) For easy identification, labeling DNA stained with fluorescent material at the end of the ss-DNA 10 to be analyzed can be conjugated (ligation) using a lyase. When the labeling DNA is conjugated to the end of the ss-DNA 10, the position of the ss-DNA 10 can be more easily identified, and the moving part 50 is moved to probe the upper end of the labeling DNA 70. ) It will be attached to the end surface.

Then, a voltage is applied to the measurement electrodes 300 provided at both ends of each nanoribbon 200 by a voltage applying unit so that a predetermined current flows through the nanoribbon 200 (S300). Then, a voltage difference or a fluid pressure difference is applied between one surface and the other surface of the substrate 500 to allow the biopolymer to pass through the nanopores 100 (S400).

Then, a voltage is applied to the electrode element 5. When a voltage is applied to the electrode element 5, the electrode element 5 located at the upper end of the measuring tank 4 has a positive pole, and the electrode element 5 located at the lower part has a negative pole. Therefore, a potential difference is formed between both ends of the substrate 500 such that the ss-DNA 10 attached to the end surface of the probe 70 passes through the nanopores 100 (S400).

The control means moves the ss-DNA 100 attached to the probe 70 in the longitudinal direction of the nanopores while controlling the piezoelectric element 60 and the moving unit 50 (S500).

In addition, the unit molecules constituting the biopolymer interact with each nanoribbon 200 to change a current flowing through the nanoribbon 200 (S600). At this time, most of the current flowing through the nanoribbons 200 corresponds to an edge current. In addition, the current flowing through the nanoribbons 200 is sensed by the measuring electrodes 300 provided at both ends of the nanoribbons 200. In this case, the nanoribbons 200 are stacked on the substrate 500 with four or more insulating layers 400 interposed therebetween, and each of the nanoribbons 200 has different units at positions where the nanopores 100 are formed. Comprising a unit molecule coating unit coated with a molecule, the complementary bonding of the unit molecule and the base coating portion passing through the nanopores 100 can be induced to maximize the current change flowing through the nanoribbons 200.

In addition, the analyzing means obtains the current change data by the unit molecules forming the biopolymers passing through the nanopores 100 (S700). In addition, the analyzing means may determine the sequence of the unit molecules by combining and analyzing the obtained current change data (S800).

Claims

Nanoribbons having a specific width inducing a change in current by interaction with the closest unit molecules of the biomolecules;

Measurement electrodes formed at each of both ends of the nanoribbon in the longitudinal direction to measure the current change;

An insulating layer formed on the top and bottom surfaces of the nanoribbons to electrically insulate the nanoribbons from each other; And

Real-time molecular sequence analysis system using a stacked nanoribbon, characterized in that it comprises a; nanopores penetrating the insulating layer and the nanoribbons in the vertical direction and the unit molecules forming the biopolymer.
The method of claim 1,

Further comprising a substrate for supporting the nanoribbon, the measuring electrode and the insulating layer,

The nanopores are formed by penetrating the substrate, the insulating layer and each of the nanoribbons in a vertical direction.
The method of claim 2,

The nano- ribbon is a real-time molecular sequence analysis system using a laminated nanoribbon, characterized in that four or more are laminated on the substrate with the insulating layer therebetween.
The method of claim 3, wherein

Each of the nanoribbons

With unit molecular coatings coated with different unit molecules at the position where the nanopores are formed

Real-time molecular sequence analysis system using laminated nanoribbons, characterized in that to induce complementary bonding of the unit molecule and the coating portion passing through the nanopores.
The method of claim 1,

The nanoribbons include zig-zag graphene or two-dimensional topological insulators.

The inside of the nanoribbon has a insulating (nearly insulating) and the edge is a real-time molecular sequence analysis system using a laminated nanoribbon characterized in that the conductive properties.
The method of claim 1,

The measuring electrode is made of a conductor including at least one of gold, silver, copper, platinum, palladium, titanium, nickel, and cobalt, and is electrically connected to the nanoribbon. Molecular Sequence Analysis System.
The method of claim 1,

The insulating layer is formed of an electrical insulator including at least one of a silicon oxide film, a silicon nitride film, an aluminum oxide film, a silver oxide film, a zinc oxide film, and a hafnium oxide film, and insulates the nanorib from an electrolyte and other nanoribbons. Real-time molecular sequence analysis system using a laminated nanoribbon, characterized in that.
The method of claim 1

At the edge of the nanoribbon where the nanopores are formed,

A real-time molecular sequence analysis system using laminated nanoribbons, characterized in that for bonding a hydrogen atom or a nitrogen atom to a carbon atom having a dangling bond.
The method of claim 1,

Real-time molecular sequence analysis system using the laminated nanoribbon, characterized in that the thickness of the nanoribbon is 5Å or less.
The method of claim 1,

Real-time molecular sequence analysis system using laminated nanoribbons, characterized in that the diameter of the nanopores is less than 10nm.
The method of claim 1,

Real-time molecular sequence analysis system using a stacked nanoribbon, characterized in that it further comprises a voltage applying unit for applying a voltage to the measuring electrode.
The method of claim 11,

Acquiring current change data based on the current measured by the measuring electrode, and analyzing means for identifying the sequence of the unit molecules of the biopolymer based on the current change data using a stacked nanoribbon Real time molecular sequence analysis system.
The method of claim 1,

The substrate is a separation wall that separates both ends of the measuring tank in which the electrolyte is maintained to maintain the biopolymer therein,

An electrode element disposed at each of both sides of the substrate to be spaced apart from each other at a predetermined interval to form a potential difference at both ends of the substrate to supply a force to move the biopolymer through the nanopores; And

A probe having one end surface coupled to one end of the biopolymer, and having one side connected to the other side of the probe and having a piezoelectric element tensioned or compressed by an applied current, the biopolymer passing through the nanopores Real-time molecular sequence analysis system using laminated nanoribbons, characterized in that to control the.
The method of claim 13,

A moving part connected to the piezoelectric element to move the piezoelectric element and the probe integrally in x, y and z axis directions; And

Real-time molecular sequence analysis system using the stacked nanoribbon further comprising a control means for controlling the moving unit and the piezoelectric element.
Applying a voltage to a measurement electrode connected to both longitudinal ends of each nanoribbon such that a current flows through the nanoribbon;

Applying a voltage difference or a fluid pressure difference between one side and the other side of the substrate to allow the biopolymer to pass through the nanopores;

Changing the current flowing through the nanoribbons by interacting with the nanoribbons unit molecules forming the biopolymer;

Sensing a change in current flowing through the nanoribbon; And

Real-time molecular sequence analysis method using the stacked nanoribbons, comprising the step of: combining and analyzing the current changes flowing through each of the nanoribbons to determine the sequence of the unit molecules.
The method of claim 15,

The current flowing through the nanoribbon is a real-time molecular sequence analysis method using a stacked nanoribbon, characterized in that it comprises an edge current (edge current).
The method of claim 15,

The step of changing the current

The nanoribbons are laminated on a substrate with four or more insulating layers interposed therebetween, and each of the nanoribbons includes unit molecular coatings coated with different unit molecules at positions where the nanopores are formed.

Real-time molecular sequence analysis method using a laminated nanoribbon, characterized in that the complementary bond between the unit molecule and the coating portion passing through the nanopore is induced to change the current flowing through the nanoribbon.
The method of claim 15,

The detecting step and the determining step

Based on the change in current flowing through the nanoribbons sensed by the measuring electrode, the analysis means combines and analyzes the change in current flowing through the nanoribbon to determine the sequence of unit molecules. Molecular sequence analysis method.
The method of claim 15,

Driving the moving part to position the probe close to the nanopores;

Applying a voltage to an electrode element so as to form a potential difference at both ends with respect to a substrate partitioning the measurement tank, wherein the biopolymer passes through the nanopores; And

And controlling the piezoelectric element and the moving unit of the moving means to adjust the moving speed of the biopolymer and to move the biopolymer.