JP2011045944A - Nanoribbon and manufacturing method thereof, fet using nanoribbon and manufacturing method thereof, and base sequence determination method using nanoribbon and apparatus for the same - Google Patents

Abstract

The present invention realizes sequencing of DNA and the like at high speed and without any limitation on the sequence length and with a simple configuration.
SOLUTION: The ssDNA 415 contained in one reservoir 412 is moved through the nanochannel 411 while being stretched by applying an electrophoretic electric field. Each base of ssDNA interacts with the active material in the nanoribbons 401 to 403 having a sandwich structure placed in the nanochannel, so that a detection signal that varies depending on the type of base is obtained. From this detection signal, the base sequence on the ssDNA is identified.
[Selection] Figure 4

Description

  The present invention relates to a nanoribbon and a method for producing the same, an FET using the nanoribbon and a method for producing the same, and a method and apparatus for analyzing molecules, particularly nucleic acids, using the nanoribbon. The nanoribbon of the present invention is made of a single layer, a double layer, several layers, or a multilayer material such as graphene. In addition, the method and apparatus using the nanoribbon of the present invention, in particular, stretches and drives a DNA or RNA molecule in a nanochannel and drives such DNA or RNA molecule in a controlled manner in the active region of the nanoribbon. Sequencing directly in real time for each base by interacting with. Gene sequence information is obtained by detecting base-specific interactions for each individual base between the nucleotide base and the edge of the nanoribbon. This interaction occurs when the nucleic acid molecule passes the edge of the nanoribbon.

Taking deoxyribonucleic acid (DNA) as an example, a gene in an organism is composed of DNA in which four types of organic bases are arranged in pairs along a double helix structure. Here, the four types of bases are adenine (A), guanine (G), cytosine (C) and thymine (T). In order to obtain specific genetic information, it is essential to determine the specific order of base pairs along this structure.
Sanger DNA sequencing technology has been the mainstay of the sequencing industry for about 30 years. Even today, Sanger DNA sequencing is a reliable and accurate method for sequencing and can read up to 99.999% of base pairs in lengths of 1000 base pairs or more. However, the Sanger sequencing method relies on chemical methods to read the bases G, C, A, and T in DNA, and for the purpose of reading individual genetic information, it is a great improvement on this technology. Nevertheless, it is still slow and expensive. In this method, it takes about 10 million to 25 million US dollars for determination of a single human gene sequence, and about 20,000 to 50,000 US dollars for determination of a bacterial gene sequence.
For example, as explained in Non-Patent Document 1, several other methods for DNA sequencing have been developed over the past few years with the aim of reducing costs and increasing sequencing speed. These development results have led to a significant reduction in sequencing costs and at the same time increased DNA sequencing speed. However, these novel sequencing techniques exhibit various reading lengths, various error rates, and various error profiles, both compared to previous data and compared to each other (see Non-Patent Document 2). See This has a great impact on the application of these technologies.
There is a strong demand for the development of new DNA sequencing technologies, and new technologies such as nanopore sequencing technologies have been developed. The principle on which nanopore sequencing technology is based is that single-strand DNA (ssDNA) molecules are driven by electrophoresis, and nanoscale pores (1.5-5 nm) are strictly in series. (See Non-Patent Document 3). Changes in electrical signals such as ionic current block, transverse tunneling current, capacitance, etc. are recorded to identify DNA sequences. However, this technique does control the speed and direction of DNA while passing through the nanopore, that nanoparticles can be trapped in the pore, large noise in the electrical signal, embedded and accurate Great challenges remain, such as creating suitable nanopores with electrically conductive and robust probe pairs aligned to each other.
Specifically, US Patent Publication No. 2006/024497 cited as Patent Document 1 discloses individual nanowires and nanotubes having a diameter of less than 0.7 nm, such as single-walled carbon nanotubes (SWCNT). A single molecule sequencing method based on a nanotransistor fabricated by using the method is disclosed. In this method, different currents are measured for each base according to the charge transfer between the DNA molecule and the SWCNT. In order to obtain single base resolution, this method requires the nanowire or nanotube diameter to be less than 0.7 nm. Significant difficulties in this method are rooted in how to make such nanowire / nanotube nanotransistors. Both carbon nanotubes and other nanowires / nanotubes are provided as having various sizes and structures, and their electrical properties depend on chirality and diameter. Currently, nanostructures such as a single type of carbon nanotubes can be manufactured to easily position individual nanowires / nanotubes for the elemental processes required for the DNA sequencing device in question. Is impossible. Therefore, an improved system and feasible method for inexpensive and fast DNA sequencing is desired.

  It is an object of the present invention to provide a novel method and apparatus that improves upon the prior art described above for sequencing. More specifically, an object of the present invention is to realize sequencing of DNA and the like at high speed and without any limitation on the sequence length. It is also an object of the present invention to provide a nanoribbon and nanoFET used for such sequencing and a method for producing the same.

In sequencing detectors according to one aspect of the invention, the detection mechanism is based on base-specific interactions with the active region of the nanoribbon. Such nanoribbons may be incorporated into field-effect nano-transistor (NanoFET) structures. For example, the channel conductance of a nanoFET changes when the nanoFET interacts with a single base on single stranded DNA. This change can be further modulated by the gate electrode field. It has been experimentally shown that nanostructure-based biosensors can detect base mismatches in DNA molecules (see Non-Patent Document 4). This result suggests the possibility of sequencing DNA by detecting base-specific interactions using nanotransistors. Furthermore, a base-specific interaction with graphene has been shown theoretically (see Non-Patent Document 5).
This detector for sequencing includes nanoribbons or electric fields as electrical or optical detectors made of single-layer, double-layer, several-layer (3-10 layers) or multilayer (more than 10 layers) materials. An effect transistor (FET) and a nanofluidic system are provided. It is an important matter of the present invention to use a layered material of monolayer, bilayer, several layers or multiple layers such as nanoribbon of graphene (thickness of about 0.335 nm) as an active member of nanoribbon or nanoFET, This achieves resolution corresponding to the individual bases. The base-specific interaction does not take place at the atomic plane of the active material sheet, but is a monolayer, bilayer, several layers or multi-layered material, for example, the cross section of an active material sheet such as a graphene sheet is the surface of the nanoribbon (In this application, this cross section of the active material sheet sandwiched between the insulating materials that appears on the surface of the nanoribbon is referred to as “the active region of the nanoribbon”).
According to another aspect of the present invention, there is provided a nanoribbon having a structure in which a material which may be a single layer or a plurality of layers, for example, graphene, is sandwiched between insulators, and a method for manufacturing the nanoribbon. Here, the material of the single or multiple layers appears on the surface of the nanoribbon as a cross-section with a thickness ranging from 0.1 nm to 10 nm (ie, as the active region of the nanoribbon).
According to another aspect of the present invention, a simple method for manufacturing a nanoFET using the edge of such a nanoribbon as an active material is provided.
Reliable detection relies on precise control of the movement of DNA in the nanochannel and the interaction of the DNA with the edge of the nanoribbon or the active material of the nanoFET. According to yet another aspect of the present invention, such dynamic control of motion is achieved by using programmable and mutually perpendicular electric fields. Such an electric field is, for example, an electrophoretic electric field applied between the electrodes provided in each reservoir, and a holding electric field applied between the gate electrode and an electrode below or in the nanochannel.
According to yet another aspect of the present invention, the electrophoretic field is driven by linearly stretching single-stranded DNA (and of course for other nucleic acids). An appropriately polar holding field can direct the negatively charged phosphate chain away from the active region of the nanoribbon and direct its nucleotides to interact in close proximity to the active region of the nanoribbon for base discrimination. The holding field also prevents single-stranded DNA from drifting during the measurement. Thus, the dynamics of motion in the nanochannel is precisely controlled by the application time and amplitude of these electric fields.
According to yet another aspect of the invention, the repulsive electric field between the nanochannel sidewalls is used to reject negatively charged single-stranded DNA from the nanochannel walls.
When a single-stranded DNA molecule is moved through a nanochannel, it can be placed on the edge of the active material of a nanoribbon or nanoFET, for example, under a holding pulse electric field so that the current between the source and drain electrodes can be measured. Each base contacts (the nano-FET shown in FIG. 3 has the surface of the nanoribbon covered with an insulating layer, so strictly speaking, in this case, the base is not in direct contact with the active region of the nanoribbon. However, it is referred to herein as “contact”, including close enough to cause a detectable interaction). This current between the source and drain electrodes varies from base to base due to base-specific interactions with the active material. Furthermore, nanoribbons or nanoFET arrays can be easily integrated in parallel or directly for repeated measurement and comparison. In principle, this sequencing method has the advantages of unlimited reading length and simple preparation of the material and high speed sequencing.

  According to the present invention, nucleobase sequencing can be performed at high speed and without limitation on the sequence length. Also, nanoribbons and nanoFETs that can be used for such sequencing can be easily created.

It is a figure explaining the method of processing a graphene nano ribbon, and the structure of such a nano ribbon according to this invention. It is an AFM (atomic force microscope) image (a) and Auger electron spectrum (b) of graphene on the nickel surface. It is a figure which shows the example of the method of manufacturing nanoFET using nanoribbon, (a) Transfers graphene on a substrate, (b) Patterns a source electrode and a drain electrode, (c) Provides an insulating layer, (D) shows a step of patterning the gate electrode. 1 is a diagram illustrating an example of a nanoFET DNA sequencing device according to the present invention in which a nanofluidic system is integrated into a nanoFET array. FIG. FIG. 6 shows another nanoFET DNA sequencing device according to the present invention, where a nanoFET is created on top of a nanofluidic system. FIG. 4 shows another nanoFET DNA sequencing device according to the present invention in which nanofluidic nanochannels are incorporated into a nanoFET. FIG. 3 shows another nanoFET DNA sequencing device according to the present invention that uses optical techniques to detect base-specific interactions with nanoribbon structures. FIG. 2 shows another nanoFET DNA sequencing device according to the present invention that uses optical technology to detect base-specific interactions with a nanoribbon structure, where an optical signal is biased by a bias on a two-terminal electrode. Can be increased. FIG. 2 shows another nanoFET DNA sequencing device according to the present invention that uses optical techniques to detect base-specific interactions with a nanoribbon structure, where bias is applied to the source, drain, and gate electrodes. The optical signal can be increased. Diagram illustrating DNA sequence identification by programmable and mutually perpendicular electric field synchronization and coordination and interaction detection for DNA drive and transfer dynamics, and detection of base-specific interactions with the active region of the sandwich structure It is.

The foregoing summary, as well as the following detailed description of the embodiments, will be better understood with reference to the following drawings. For the purpose of illustrating the invention, the following description and drawings are presented as presently preferred. However, it should be understood that the invention is not limited to the specific methods, configurations, apparatus, and exact arrangements disclosed herein. Furthermore, it should be noted that these drawings are not necessarily drawn to scale.
The present invention relates to an apparatus and method for performing sequencing of nucleic acids (DNA or RNA) on a single molecule. More specifically, the present invention relates to obtaining gene sequence information by directly reading DNA molecules or RNA molecules for each base. In the following description and drawings, there is often a description that only one technique or mechanism according to the present invention is used for easy understanding. However, it should be noted that it is possible and preferred to use the technique repeatedly and to use multiple mechanisms unless explicitly stated.
In the following, an apparatus and method for sequencing single molecule DNA or RNA is described. Such devices have nanoribbons or nanoFETs for analytical sensors, nanofluidic mechanisms for linearization and drive of individual DNA or RNA molecules, and programmable gradient electric fields for control of DNA or RNA movement dynamics. is doing.
Considering the size of nucleotides and the separation of nucleotides in stretched or unstretched DNA molecules, the nanoribbons of the present invention can be single layer, double layer, several layers (3-10 layers), or multiple layers (from 10 layers). Even many layers are created using materials). The material of the layer is not limited to this, for example, graphite, superlattice MoS ₂ , NbSe ₂ , Bi ₂ Sr ₂ CaCu ₂ O _x , boron nitride, dichalcogenides, trichalcogenides, Tetrachalcogenides materials, where the thickness of the layered material is in the range of 0.1 nm to 10 nm, in particular in the range of approximately 0.1 nm to 2 nm, more preferably in the range of approximately 0.1 nm to 1 nm. Preferably, the layered material is single layer graphene, double layer graphene, or several layers (3-10 layers) graphene. Graphene, a two-dimensional honeycomb lattice of sp ² bonded carbon atoms, is a single layer graphite with a theoretical thickness of about 0.335 nm. Furthermore, the present invention provides a new and simple method for making nanoribbons comprising single-layer, bilayer, several-layer or multi-layer layered materials, and the structure of nanoFETs using such nanoribbons.
In FIG. 1, a modified or unmodified graphene sheet 2 synthesized by various methods such as mechanical exfoliation, graphene oxide, and chemical vapor deposition (CVD) is used as SiO ₂ . It is transferred onto an insulating substrate 3 and then covered with another insulating layer 1 such as a polymer. There is virtually no limit to the thickness of these insulating layers. It should be noted here that it is relatively easy to handle such a thicker sandwich structure. This sandwich structure is processed into a ribbon having a shape cut in a direction substantially perpendicular to the surface of the graphene sheet 2 by various techniques such as lithography, focused ion beam, reactive ion etching, and plasma etching. The nanoribbon thus produced has a shape in which a very thin cross section of the graphene sheet 2 appears in the center, and there are portions made of an insulator on both sides of the cross section. Note that there is no limit on the width of this ribbon. The ribbon has a wide range of 100 nm to 2 mm, so it is easy to handle and position the ribbon. Such a sandwich structure enables the nanoribbon active region to reliably sequence long DNA strands. Of course, instead of the graphene sheet, a layer of a material exemplified above or other than that may be used.
FIG. 2 shows an AFM image (a) and Auger electron spectrum (b) of graphene grown on a Ni surface using CVD techniques. The reason why two Auger electron spectra are shown in FIG. 2B is that the results of measurement at two different locations on the sample are shown.
Using this technique, it is possible to easily obtain very large free-standing graphene by etching the Ni layer and transfer it onto any kind of substrate such as SiO ₂ . The transfer destination substrate is preferably a flat substrate at the atomic level in order to reduce defects that may cause graphene.
FIG. 3 shows an example of the basic steps for manufacturing a field effect transistor using nanoribbons. This manufacturing step includes (a) transferring the nanoribbon onto the substrate 4, (b) patterning the source electrode 5 and the drain electrode 6, (c) creating the insulating layer 7, and (d) Patterning the gate electrode 8.
The sequencing mechanism of the present invention is not the atomic facet of monolayer, bilayer, several layers or multilayer layered materials such as graphene, but base-specific interactions between the ends of such nanoribbons and nucleotide bases. It is in. It has been observed that charge transfer along DNA molecules promotes oxidation or reduction chemical reactions (see Non-Patent Document 6). This phenomenon implies that charge transfer can occur between the DNA base and other materials such as graphene. It has been experimentally shown that nanostructure-based biosensors can detect base mismatches in DNA molecules (see Non-Patent Document 4). This result suggests that DNA sequencing can be performed electrically by detecting conductance changes using nanotransistors. Furthermore, the interaction unique to nucleotide bases with graphene has been shown theoretically (see Non-Patent Document 5). Thus, the sequence in the nucleic acid molecule can be determined by detecting the difference in electrical or optical signal for each nucleotide base that interacts with the nanoribbon in the nanochannel.

[Example 1]
FIG. 4 shows a single molecule sequencing apparatus according to the first embodiment of the present invention. This example includes nanoFETs and nanofluidic systems. NanoFETs include a substrate 404, a gate electrode 408, a dielectric layer 407, nanoribbon structures 401, 402, 403 such as sandwiched nanoribbon active regions, a source electrode 405, a drain electrode 406, and individual nanoFETs and planarization layers. an insulating layer 416 for separating the layer). Note that the functional materials for these nanoFET components are not limited in width or thickness. At this stage, the nanoribbon width can be reduced by various techniques if necessary, and the active state of the nanoribbon can be varied by various techniques such as O ₂ and hydrogen plasma, NH ₃ or other chemicals. Can be reconfigured or modified to have specific characteristics. The nanofluidic system includes an insulating layer 410, a reservoir 412, a nanochannel 411, and electrophoretic electrodes 413, 414 for loading, linearizing, and feeding individual single-stranded DNA (ssDNA) molecules. The width of the nanochannel 411 is in the range of 2 nm to 500 nm, and the depth is not limited. The nanochannel 411 is made of an insulating material suitable for implementing the functions of loading, stretching (ie, linearizing), and feeding the ssDNA 415. A conductive electrode 409 is aligned on the nanochannel 411 and the nanoFET. By applying a programmable bias to the electrode 409 and the gate electrode 408, a holding electric field is generated. By providing appropriate polarity, amplitude, and duration, programmable electrophoretic and holding electric fields precisely control ssDNA kinetics in the nanochannel 411.
Since the DNA has a chain of negatively charged phosphate groups, the ssDNA moves toward the anode under the influence of the electrophoretic field. The distance of the ssDNA step is controlled by the duration and amplitude of the electrophoresis pulse. A holding electric field of appropriate magnitude and polarity ensures that the ssDNA molecule is directed toward the nanoFET and the negatively charged phosphate group is further away from the nanoFET as desired for base detection. It can be kept facing. The holding electric field can also prevent DNA molecules from drifting during detection. Both fields are appropriately programmed to drive delivery and detection to provide sufficient time and stability for the nanoFET to detect individual bases. In the preferred practice of the invention, the operation and interaction and detection processes of these two electric fields are synchronized and coordinated to achieve extremely fast sequencing. As the ssDNA molecule moves through the nanochannel, it can contact the active region of the graphene nanoribbon one molecule at a time by the holding pulse electric field and measure the current between the source and drain electrodes To. This current varies between the various bases due to base-specific interactions with the active region of the nanoribbon. Furthermore, for repeated measurements and comparisons, multiple graphene nanoribbon nanotransistor arrays can be easily integrated into a nanofluidic system in parallel and / or in series.
[Example 2]
FIG. 5 shows another embodiment of the sequencing apparatus of the present invention. In the embodiment of FIG. 5, nanoFETs are integrated on a nanofluidic system formed on a substrate 504. The nanofluidic system includes an insulating layer 510, a reservoir 512, a nanochannel 511, and electrophoretic electrodes 513, 514 for loading, linearizing and delivering individual ssDNA 515 molecules. In order to achieve an effective interaction between the DNA base and the edge of the nanoribbon, the depth of the nanochannel 511 is preferably less than 20 nm. The nanoFET has a gate electrode 508, a dielectric layer 507, an active nanoribbon structure, that is, nanoribbons 501, 502, 503, a source electrode 505 and a drain electrode 506. Conductive electrodes 509 are aligned directly under these nanochannels and nanoFETs. By applying a programmable bias to the electrode 509 and the gate electrode 508, a holding electric field is generated. An electrophoretic electric field is generated by applying a bias between the electrophoretic electrodes 513 and 514. It should be noted that elements for separation, encapsulation, etc. are omitted in this embodiment.
[Example 3]
FIG. 6 shows yet another embodiment of the sequencing apparatus of the present invention. As shown in FIG. 6, the nanochannel 611 may be formed so that the whole or a part of the nanochannel 611 penetrates the nanoFET. The nanoFET is a nanoribbon structure 601, 602, 603, a substrate 604, a source electrode 605, a drain electrode 606, a dielectric layer 607, a gate electrode 608, and an insulating layer for separating individual nanoFETs. 616. The nanofluidic system includes a reservoir 612, a nanochannel 611, and electrophoresis electrodes 613 and 614. The insulating layer 610 is on the nanochannel 611, and a conductive electrode 609 is provided thereon. A gradient electric field is generated between the electrodes 613 and 614 and between the conductive electrode 609 and the gate electrode 608. These gradient fields are used to control the dynamics of ssDNA motion and interaction with the nanoFET. In this configuration, the interaction region between the DNA base and the nanoribbon active region is large, and therefore the influence of the base direction on the detection signal is eliminated.
[Example 4]
FIG. 7 shows sandwich-type nanoribbon structures 701 and 702 by an optical detection system using an optical technique such as energy transfer spectroscopy, absorption, and emission-emission spectroscopy. A sequencing device is provided that detects base-specific interactions with the 703 edge. The sequencing device is made on a substrate 704. Sandwich-type nanoribbon structures are integrated on a nanofluidic system. The nanofluidic system has an insulating layer 706, a reservoir 707, a nanochannel 708, and electrophoresis electrodes 709 and 710 for loading, linearizing, and feeding individual single-stranded DNA 711 molecules. In order to obtain efficient interaction between the DNA base and the edge of the nanoribbon so that it can be detected by the optical detection system 713, the nanochannel depth is preferably less than 20 nm. Conductive electrodes 705 are aligned under the nanochannels and nanoribbons. By applying a programmable bias to the electrode 705 and the electrode 712, a holding electric field is generated. The electrophoretic field is generated by applying a bias between electrodes 709 and 710.
[Example 5]
In yet another embodiment, as shown in FIG. 8, the optical signal can be increased by electrical stimulation by applying a bias between electrodes 814 and 815. Sandwich-type nanoribbon structures are integrated on the nanochannel. The nanofluidic system has an insulating layer 805, a reservoir 807, a nanochannel 808, and electrophoretic electrodes 809, 810 for loading, linearizing, and feeding individual ssDNA811 molecules. In order to obtain efficient interaction between the DNA base and the active region of the nanoribbon so that it can be detected by the light detection system 813, the nanochannel depth is preferably less than 20 nm. The conductive electrode 805 is aligned directly under the nanochannel and nanoFET. By applying a programmable bias to electrode 805 and electrode 812, a holding electric field is generated. The electrophoretic field is generated by applying a bias between the electrophoretic electrodes 713 and 714. Such a stimulus may be other stimuli such as a magnetic field in addition to the above electric field.
[Example 6]
In yet another embodiment, such as that shown in FIG. 9, the optical signal is amplified by applying a bias between the source electrode 914 and the drain electrode 915 and then modulating the gate signal via the gate electrode 912. Can do. This sequence analyzer is created on a substrate 904. NanoFETs are integrated on top of nanofluidic systems. The nanoFET includes sandwich nanoribbon structures 901, 902, and 903, a source electrode 914, a drain electrode 915, an insulating layer 915, and a gate electrode 912. The nanofluidic system includes an insulating layer 906, a reservoir 907, a nanochannel 908, and electrophoretic electrodes 909, 910 for loading, linearizing, and feeding individual ssDNA 911 molecules. In order to obtain an efficient interaction between the DNA base and the active region of the nanoribbon so that it can be detected by the light detection system 913, the nanochannel depth is preferably less than 20 nm. The conductive electrode 905 is aligned directly below the nanochannel and nanoribbon. By applying a programmable bias to electrode 905 and electrode 912, a holding electric field is generated. By applying a bias between electrophoretic electrodes 909 and 910, an electrophoretic electric field is generated.
In one aspect of the embodiment, the repulsive electric field between the side walls of the nanochannel is used to reject negatively charged ssDNA from the sidewalls of the nanochannel. The nanochannel can be linear, S-shaped, branched, or any other shape. In one aspect of the embodiment, a transparent material is used for the convenience of signal detection.
In order to achieve a cooperative operation process, the base detection process in the sequencing apparatus described above is synchronized with the programmed electrophoretic and holding electric field operations, as shown in FIG. FIG. 10 shows an electrophoresis pulse and a holding pulse for generating an electrophoretic electric field and a holding electric field, an interaction detection timing signal that is a timing for detecting an interaction between the base and the nanoribbon active region, and a base-specific mutual signal. The action detection results are illustrated with the time axes aligned. In addition, identification results (A, G, C, T) of individual bases are also shown on the detection results of base-specific interactions.
In FIG. 10, the DNA is stepped by one base at the timing E1, E2,..., En of the electrophoresis pulse, and thereafter, the timing H1, H2,. Then, the DNA that has advanced is drifting, that is, kept so as not to move any further. While the DNA thus advanced is held, the base-specific interaction between the base on the DNA and the nanoribbon active region is detected at timings D1, D2,..., Dn. Specifically, this detection is performed by measuring a detection output (a source current or drain current of a nano-FET in Examples 1 to 3 and an output of an optical detection system in Examples 4 to 6) with a circuit (not shown). Is done. By using computerized system control and data acquisition, the characteristic profile of the interaction signal for each of the four different DNA bases can be determined for all or any of the detection techniques by referring to the nucleotides of a known DNA molecule. It can be determined by any combination. These feature signal profiles can be used to identify DNA base sequences.
In order to achieve the optimization of the base identification performance, a plurality of detectors based on sandwich-like nanostructures as described above are integrated in series and / or in parallel with the nanofluidic system. An improvement in accuracy can be achieved by mutual comparison and correction.
While the above examples are for DNA sequencing, it will be appreciated that the present invention encompasses other types of nucleic acid sequencing.
While the presently preferred embodiments of the invention have been disclosed and described above, various changes and modifications may be made by those skilled in the art without departing from the scope of the invention as defined in the claims. It will be clear that this is possible.

  Since the present invention can realize sequencing of DNA molecules etc. at a very high speed and without any restriction on the length, it can be used in a wide range of industrial fields.

US Published Patent 2006/0246497

J. et al. Shendure, H.M. L. Ji, Next-generation DNA sequencing. Nature Biotechnol. 26, 1135-1145 (2008) M.M. Pop, S.M. L. Salzberg, Bioinformatics challenges of new sequencing technology. Trends in Genetics 24, 142-149 (2008) Branton, D.C. , Et al. The potential and challenges of nanosequencing. Nature Biotechnol. 26, 1146-1153 (2008) Hahm, J .; & Lieber, C.I. M.M. Direct ultrasensitive electrical detection of DNA and DNA sequence variations using nanowires nanosensors. Nano Lett. 4, 51-54 (2004) Antony, J.M. & Grimme, S .; Structures and interaction energies of stacked graphene-nucleobase complexes. Phys. Chem. Phys. Chem. 10, 2722-2729 (2008). Varghese, N.A. , Et al. Binding of DNA nucleobases and nucleosides with grapheme. Chem. Phys. Chem. 19, 206-210 (2009) Elias, B.M. , Shao, F .; & Barton, J.A. K. Charge migration along the DNA duplex: hole versus electron transport. J. et al. Am. Chem. Soc. 130, 1152-1153 (2008)

DESCRIPTION OF SYMBOLS 1 Insulating layer 2 Graphene sheet 3 Insulating substrate 4 Substrate 5 Source electrode 6 Drain electrode 7 Insulating layer 8 Gate electrode 401, 402, 403, 501, 502, 503, 601, 602, 603, 701, 702, 703, 801, 802, 803, 901, 902, 903 Nano ribbon structure 404, 504, 604, 704, 804, 904 Substrate 405, 505, 605, 914 Source electrode 406, 506, 606, 915 Drain electrode 407, 507, 607, 916 Dielectric Layers 408, 508, 608, 912 Gate electrodes 409, 509, 609, 705, 712, 805, 812, 814, 815, 905 Electrodes 410, 416, 510, 610, 616, 706, 806, 906 Insulating layers 411, 511 611, 708, 808, 908 Channels 412, 512, 612, 707, 807, 907 Reservoirs 413, 414, 513, 514, 613, 614, 709, 710, 809, 810, 909, 910 Electrophoretic electrodes 415, 515, 615, 711, 811, 911 ssDNA
713, 813, 913 Optical detection system

Claims (29)

A nanoribbon having the following features (a) to (c).
(A) A first part is provided.
(B) A second portion and a third portion that have insulating properties and sandwich the first portion therebetween are provided.
(C) The first portion is a single layer or a plurality of layers, and the cross section of the single layer or the plurality of layers is formed on the surface of the nanoribbon with a width of 0.1 nm to 10 nm. Appears as a part. The first portion is a layered or multilayer material selected from the group consisting of graphite, superlattice MoS ₂ , NbSe ₂ , Bi ₂ Sr ₂ CaCu ₂ O _x , boron nitride, dichalcogenide, trichalcogenide, and tetrachalcogenide. The nanoribbon according to claim 1. The nanoribbon according to claim 2, wherein the first portion is a sheet of the layered or multilayer material having 1 to 10 layers. The nanoribbon according to claim 3, wherein the first portion is a 1 to 10 layer graphene sheet. The nanoribbon according to any one of claims 1 to 4, wherein a width of the nanoribbon is in a range of 100 nm to 2 mm. The nanoribbon manufacturing method provided with the following steps (a) to (d).
(A) A sheet of material having a cross-sectional width in the range of 0.1 nm to 10 nm is prepared.
(B) The sheet is disposed on the first insulating layer.
(C) Cover the sheet with a second insulating layer.
(D) The nanoribbon having the first insulating layer, the sheet, and the second insulating layer, and the sheet sandwiched between the first insulating layer and the second insulating layer on a surface thereof A nanoribbon with a cross section appears. The sheet, graphite, superlattice _{MoS 2, NbSe 2, Bi 2} Sr 2 CaCu 2 O x, boron nitride, dichalcogenides, Torikarukogenido is selected from the group consisting of tetra chalcogenide, nanoribbons method according to claim 6, wherein . The nanoribbon manufacturing method according to claim 6 or 7, wherein the sheet is a graphene sheet having 1 to 10 layers. The nanoribbon manufacturing method according to any one of claims 6 to 8, wherein a width of the nanoribbon is in a range of 100 m to 2 mm. NanoFET provided with the following (a) to (e).
(A) The nanoribbon according to any one of claims 1 to 5, which is provided on a substrate.
(B) A source electrode placed in contact with one end of the nanoribbon.
(C) A drain electrode installed to come into contact with the other end of the nanoribbon.
(D) A dielectric layer installed so as to cover the nanoribbons.
(E) A gate electrode disposed on the dielectric layer. A nano-FET manufacturing method provided with the steps (a) to (e).
(A) The nanoribbon according to any one of claims 6 to 9 is prepared.
(B) The nanoribbon is transferred to a substrate.
(C) A source electrode and a drain electrode are patterned so as to be in contact with each end of the nanoribbon.
(D) A dielectric layer covering the nanoribbon is provided.
(E) A gate electrode is patterned on the dielectric layer. A nucleic acid sequencing apparatus provided with the following (a) and (b).
(A) The nanoribbon according to any one of claims 1 to 5.
(B) moving the nucleic acid therein to provide a detection signal for identifying the individual base of the nucleic acid by moving the nucleic acid across the cross section appearing on the surface of the nanoribbon and interacting with the cross section; Nanofluid system. The device according to claim 12, wherein the nanofluidic system includes the following (b-1) and (b-2).
(B-1) A nanochannel through which the nucleic acid moves.
(B-2) Means for applying an electric field that drives the nucleic acid by stretching it, thereby moving the nucleic acid across the cross section appearing on the surface of the nanoribbon. The apparatus according to claim 13, wherein the electric field that is driven to stretch moves the nucleic acid stepwise, thereby moving the nucleic acid by the length of one base of the nucleic acid per step. A means is further provided for holding the nucleic acid in a state in which the base of the nucleic acid is close to the cross section appearing on the nanoribbon by applying a holding electric field to the nucleic acid moving through the nanochannel. 15. The apparatus according to claim 13 or claim 14. The apparatus of claim 15, wherein the holding electric field is complementary to the stretching and driving electric field. 17. The source electrode and the drain electrode, each of which is in contact with one end of the nanoribbon to form a nano FET, are further provided, and the detection signal is obtained as a current between the source electrode and the drain electrode. The device according to any one of the above. The apparatus according to any one of claims 12 to 16, wherein an optical detection system is further provided above the cross section appearing on the surface of the nanoribbon to obtain the detection signal. The apparatus of claim 18, further comprising means for applying a stimulus to the nanoribbon to increase the detection signal. The apparatus of claim 19, wherein the stimulus is an electric or magnetic field. A nucleic acid sequencing apparatus provided with the following steps (a) to (c).
(A) A nanofluid system is provided that is driven through nucleic acids.
(B) The nanoribbon according to any one of claims 1 to 5 is provided in the nanofluid system.
(C) moving the nucleic acid through the nanofluidic system, moving the nucleic acid across the cross-section appearing on the surface of the nanoribbon to interact with the cross-section, and A detection signal for identifying the base of is generated. 23. The method of claim 21, wherein the driving step comprises applying an electric field that drives the nucleic acid to stretch and drive the nucleic acid across the cross-section on the surface of the nanoribbon and interact with the cross-section. 23. The method of claim 22, wherein the electric field driven to extend moves the nucleic acid stepwise, thereby moving the nucleic acid by the length of one base of the nucleic acid per step. Further comprising applying a holding electric field to the nucleic acid moving through the nanofluidic system to hold the nucleic acid in proximity to the cross-section where the base of the nucleic acid appears on the nanoribbon. 24. The method of claim 22 or 23. 25. The method of claim 24, wherein the holding electric field is complementary to the stretched and driving electric field. The method further comprises providing a source electrode and a drain electrode that respectively contact one end of the nanoribbon to form a nanoFET so that the detection signal is obtained as a current between the source electrode and the drain electrode. 26. A method according to any one of claims 21 to 25. 26. The method according to any one of claims 21 to 25, further comprising providing an optical detection system above the cross-section appearing on the surface of the nanoribbon to obtain the detection signal. 28. The method of claim 27, further comprising applying a stimulus to the nanoribbon to increase the detection signal. 30. The method of claim 28, wherein the stimulus is an electric or magnetic field.