JP2003021637A - Dna chip provided with cover plate - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a DNA chip capable of uniform deployment of a liquid specimen for hybridization in a hybridization region. SOLUTION: The DNA chip is provided with a hybridization implementation region containing a plurality of DNA fragment probe fixation spots and with a liquid specimen application region adjacent to the implementation region. The DNA chip also has a cover plate installed above the implementation region with a gap across which a water-based DNA fragment liquid specimen may deploy on the implementation region thanks to capillarity. A liquid specimen application aperture is provided on the boundary between the liquid specimen application region and the hybridization implementation region at a position corresponding to a location below one of the sides of the cover plate, or a cutout or an opening may be provided on a side of the cover plate positioned above the boundary region.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an assembly consisting of a DNA chip and a cover plate, which is very useful for simultaneous analysis of gene expression, mutation, polymorphism and the like.

[0002]

2. Description of the Related Art Attempts have been made not only to elucidate the gene structure of a wide variety of organisms but also to elucidate their gene functions on a genome scale, and technological development for efficiently analyzing gene functions. Is also progressing. A DNA chip (that is, a DNA microarray) is a high-density array in which a large number of DNA molecules are arrayed on a solid-phase carrier such as a slide glass, and determines the nucleotide sequence of nucleic acids, as well as gene expression, mutation and polymorphism. It is very useful for simultaneous analysis such as. Analysis of genetic information using this DNA chip
It can be applied to biomolecules other than DNA, drug discovery research,
Since it is extremely useful for diagnosing diseases and developing preventive methods, further development of DNA chip production technology and analysis system of the obtained data is desired.

When detecting a nucleic acid such as DNA using a DNA chip, for example, in the case of a labeling method, first, a radioisotope (RI) is applied to a DNA fragment (probe) densely aligned on the surface of a solid support. Or a sample nucleic acid fragment labeled with fluorescence (target nucleic acid) is hybridized. At this time, molecules having a base sequence complementary to the probe DNA in the target nucleic acid are
Complementarily binds (hybridizes) with A. Then, the signal of this hybridized labeled target molecule is measured to identify the other hybridizing probe molecule. Specifically, the radiation intensity or fluorescence intensity on the chip is measured by RI or a fluorescence image scanner, and the obtained image data is analyzed. Further, in the case of the electrochemical method, instead of labeling the sample nucleic acid fragment, a potential is applied to the DNA chip in the presence of the electrochemically active threading type intercalator to flow between the intercalator and the chip. The signal is measured by measuring the value of the electric current. Therefore, according to the DNA chip, thousands to tens of thousands of molecules can be simultaneously reacted and detected, and a large amount of gene information can be obtained in a short time with a small amount of sample.

[0004]

Heretofore, hybridization of a sample nucleic acid fragment with a DNA fragment on a DNA chip has generally been carried out by dropping an aqueous sample solution containing the sample nucleic acid fragment onto a planar DNA chip, It is carried out by placing a cover plate in the shape of a circle on the chip so that the sample solution spreads on the surface of the chip and bringing the sample nucleic acid fragment and the DNA fragment into contact with each other. However, generally, DNA fragments are fixed as hundreds to tens of thousands of spots on a DNA chip having a length and width of several centimeters, and in reality, the sample solution is sufficiently spread over each spot. It was difficult to spread the sample solution evenly over the entire surface.

An object of the present invention is to provide a DNA chip assembly capable of uniformly spreading a sample solution for hybridization on the hybridization region on the surface of the DNA chip.

[0006]

As a result of studying the nonuniformity of the spread of the sample solution during hybridization, the present inventor found that a spacer was placed between the DNA chip and the cover plate, and It was found that the sample liquid after spotting can be uniformly spread over the entire hybridization region by the capillary phenomenon by forming a minute gap between them when the cover plates are superposed on each other. Further, by further providing a minute recess for dropping the sample solution on the DNA chip, or by providing a cutout portion or an opening portion on the side edge portion of the cover plate, the efficiency can be improved with the cover plate installed. Good and uniform sample solution DN
It has been found that it is possible to develop in the hybridization area on the A chip.

[0007] Therefore, the present invention provides a DNA chip having an hybridization surface including a plurality of DNA fragment probe fixing spots and a sample solution spotting area disposed in contact with the area, and the DNA chip. In a DNA chip with a cover plate, which comprises a cover plate provided above a hybridization execution region of the chip with a gap that allows the aqueous DNA fragment sample liquid to spread to the hybridization region by capillary action, the sample liquid With a cover plate, characterized in that a sample solution spotting hole portion is provided on the boundary between the spotting area and the hybridization execution area and at a position corresponding to the lower side of one side of the cover plate adjacent to the boundary. D
It is on the NA chip.

The present invention also provides a DNA chip, a cover plate, and an upper surface of the hybridization execution region of the DNA chip, the upper surface of which is provided with a hybridization execution region and a sample solution spotting region disposed in contact with the region. A spacer for arranging the cover plate with an interval that allows the aqueous DNA fragment sample solution to spread to the hybridization region by the capillary phenomenon, and the boundary between the sample solution spotting region and the hybridization execution region. There is also a DNA chip assembly characterized in that a sample solution spotting hole is provided on the top.

According to the present invention, a sample solution spotting area and a hybridization execution area of a DNA chip having an upper surface having a hybridization execution area and a sample solution spotting area disposed in contact with the area. There is also a DNA chip characterized in that a sample solution spotting hole is provided on the boundary.

Further, the present invention provides a DNA chip having an upper surface with a hybridization execution region including a plurality of DNA fragment probe fixing spots and a sample solution spotting region arranged in contact with the region, and the DNA chip. In a DNA chip with a cover plate, which comprises a cover plate provided above a hybridization execution region of the chip with a gap that allows the aqueous DNA fragment sample liquid to spread to the hybridization region by capillary action, the sample liquid There is also a DNA chip with a cover plate, characterized in that one side of the cover plate located above the boundary between the spotting region and the hybridization execution region is provided with an opening or notch for spotting the sample liquid. .

Furthermore, the present invention provides a DNA chip, a cover plate, which has a hybridization execution region and a sample solution spotting region arranged in contact with the hybridization region on the upper surface.
And a spacer for arranging the cover plate above the hybridization execution region of the DNA chip via a space that allows the aqueous DNA fragment sample solution to spread to the hybridization region by capillary action. D, which is provided with an opening or notch for wearing the sample liquid spot on one side
There is also an NA chip assembly.

Further, according to the present invention, spacers are provided on both side portions, and a diameter of 100 to 200 is provided on a side other than the side portions.
There is also a cover plate having an opening or notch in the range of 0 μm.

Preferred embodiments of the present invention are as follows. (1) The cover plate is made of a transparent glass material or a resin material. (2) The distance between the DNA chip and the cover plate is 5 to 2
It is in the range of 00 μm. (3) The diameter of the opening of the hole for spotting the sample solution of the DNA chip is in the range of 100 to 2000 μm, and the depth of the hole is in the range of 5 to 200 μm. (4) The diameter of the opening or notch for wearing the sample liquid spot on the cover plate is in the range of 100 to 2000 μm, and the depth of the hole is in the range of 5 to 200 μm. (5) The sample liquid spot wearing hole is a groove-shaped hole formed along the side of the cover plate.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION The DNA chip with a cover plate or the DNA chip assembly of the present invention will be described below.
A detailed description will be given with reference to the drawings. (1) of FIG.
It is a top view which shows one aspect | mode of the DNA chip with a cover plate of this invention, (2) is sectional drawing which followed the II line in (1), and (3) is II- in (1).
It is a sectional view taken along the line II.

In FIG. 1, DNA with a cover plate
The chip is composed of a DNA chip 10 and a cover plate 15 which is removably arranged on the DNA chip 10. The DNA chip 10 comprises a planar solid phase carrier 11 and a large number of probe spots 12 fixed on the surface thereof. Each spot 12 is an aggregate of DNA fragments (probe DNA), and one end of each DNA fragment is bonded and fixed to the surface of the solid-phase carrier 11 by ionic bond or covalent bond. Further, the DNA chip 10 has one side portion exposed from the cover plate 15 and is a minute injection port for dropping the sample liquid at a position facing the center of the side edge of the cover plate 15. A recess 13 is provided.

The cover plate 15 has a concave cross-sectional shape in which the two opposing ends 16 and 17 are thicker than the central portion, and the ends (spacers) 16 and 17 are in contact with the DNA chip 10. Placed on the DNA chip 10
Between the cover plate 15 and the cover plate 15
Is occurring. At the same time, the cover plate 15 is
Although smaller than the chip 10, it is arranged so as to cover all the spots 12.

The size of the DNA chip 10 (solid phase carrier 11) is generally in the range of 76 ± 0.5 mm in length, 26 ± 0.5 mm in width, and 0.9 to 1 in thickness. ．
It is in the range of 2 mm. The size of the spot 12 is DNA
Generally, the diameter is 5 depending on the type of fragment.
It is in the range of 0 to 300 μm. The distance between the spots is generally in the range of 0 to 1.5 mm, preferably 100 to
It is in the range of 300 μm. It is desirable that the dripping sample solution does not remain in the minute recesses 13, and generally, the diameter of the opening is in the range of 100 to 2000 μm, and the depth is preferably shallow, and the range is 5 to 200 μm.

The size of the cover plate 15 is generally in the range of 22 to 50 mm in the vertical direction and 22 to 24 mm in the horizontal direction.
Is in the range. The thickness difference (height of the gap) between the end portions 16 and 17 and the central portion is generally in the range of 5 to 200 μm in order to cause a capillary phenomenon by the sample liquid,
It is preferably in the range of 10 to 50 μm.

The shape of the opening of the minute concave portion 13 is not limited to the circular shape as shown in FIG. 1A, but as shown in FIGS. The shape can be any shape such as a quadrangle (eg, trapezoid) or a hexagon.

Further, the sectional shape of the minute concave portion 13 is not limited to the quadrangular shape having a flat bottom as shown in (3) of FIG. 1, but schematic sectional views of (1) and (2) of FIG. It may be an inverted triangle or trapezoid that narrows towards the bottom, as shown in, or it may also be inclined towards the cover plate.

Further, the minute recess 13 does not necessarily have to be provided at a position facing the center of the side edge of the cover plate 15, and as shown in (1) of FIG. It may be provided at a position facing the end. Further, the number of minute recesses is not necessarily one, and may be two or more as shown in (2) and (3) of FIG.
When a plurality of minute recesses are provided, the intervals between the recesses are preferably equal. Further, the plurality of recesses may be in communication with each other, and may be formed in a groove shape along the side edge of the cover plate, for example, as shown in (4) of FIG. Alternatively, as shown in (5) of FIG. 4, concave portions are provided at two positions, that is, a position facing the side edge of the cover plate and a central portion, and these two concave portions are connected by a thin groove. May be.

FIG. 5 shows a DN with a cover plate according to the present invention.
It is a top view which shows another aspect of an A chip. In FIG. 5, the DNA chip with the cover plate is the DNA chip 2.
0 and a cover plate 25, the DNA chip 20 is composed of a planar solid phase carrier 21 and a large number of spots 22 fixed to a predetermined hybridization execution region on the surface thereof. The cover plate 25 has a concave cross-sectional shape in which two opposite end portions 26 and 27 are thicker than a central portion, and a notch portion as an injection port for dropping the sample liquid is formed in the center of one side edge portion thereof. 28 is provided. The diameter of the cutout portion 28 is generally 100 to
It is in the range of 2000 μm (when the notch is not a circle, the size approximates to a circle).

In the above embodiment, the cutout portion 28
The shape of is not limited to a semicircle, and may be any shape such as a triangle or a quadrangle (eg, trapezoid). Further, the cutout portion does not have to be provided at one place in the center of the side edge portion, but may be provided at a plurality of places, and can be provided at any position of the side edge portion. Alternatively, it may be provided in a rectangular shape or an arc shape along the side edge portion.

FIG. 6 shows a DN with a cover plate according to the present invention.
It is a top view which shows another aspect of an A chip. 6, the DNA chip with a cover plate is composed of a DNA chip 30 and a cover plate 35, and the DNA chip 30 is composed of a planar solid phase carrier 31 and a large number of spots 32 fixed on the surface thereof. Cover plate 35
Has a concave sectional shape in which two opposite ends 36, 37 are thicker than the central part. A cutout portion 38 is provided at the center of the side edge portion of the cover plate 35 as an inlet for dropping the sample solution, and the DNA chip 30 is provided on one side.
A minute recess 33 is provided at a position facing the cutout portion 38.

In the above embodiment, the cutout portion 3
It is not always necessary to provide one and one small recesses 8 and one minute recess 33, respectively, and a plurality of multiple recesses and multiple recesses 33 may be provided. In that case, the cutout portion can be provided at any position on the side edge portion. Also, the minute recesses can be provided in various positions in various shapes as described above, but it is desirable that they correspond to the cutout portions.

In the present invention, the gap for uniformly spreading the sample solution on the DNA chip by the capillary phenomenon is provided with a spacer instead of providing a spacer on the cover plate side.
It can also be provided on the DNA chip side.

FIG. 7 (1) shows a spacer-equipped D of the present invention.
It is a top view which shows another aspect of NA chip, and (2) is sectional drawing which followed the II line in (1). In FIG. 7, the DNA chip assembly is composed of a DNA chip 40 and a cover plate 45 which is detachably arranged on the DNA chip 40. The DNA chip 40 comprises a solid phase carrier 41 and a large number of spots 42 fixed on the surface thereof. The solid phase carrier 41 has a concave cross-sectional shape in which the thickness of the peripheral portion (spacer) 41a is thicker than the thickness of the central portion, and a gap 44 is formed. Each spot 42
Are fixed above this gap 44. On the other hand, the cover plate 45 is flat as in the conventional case. The cover plate 45 exposes one end of the gap 44 for facilitating the dropping of the sample solution, and covers each spot 42 so that the sample solution sufficiently spreads over each spot 42. Placed on top.

The difference in thickness (spacer thickness) between the peripheral portion 41a and the central portion of the solid phase carrier 41 is generally in the range of 5 to 200 μm, preferably in order to cause the capillary phenomenon by the sample solution. It is in the range of 10 to 50 μm.

In the spacer-attached DNA chip of the present invention, the shape of the gap of the DNA chip is not limited to the shape shown in FIG. 7, and various shapes can be used as long as an injection port for dropping the sample solution is provided. Can take the form of. For example, like the cover plate 15 in FIG.
The NA chip may have a shape in which only both end portions are thicker than the central portion. Alternatively, the minute concave portion or the cutout portion as described above may be provided separately as an injection port for dropping the sample liquid.

Each of (1) to (3) in FIG. 8 is a top view schematically showing another embodiment of the DNA chip assembly of the present invention. In (1) of FIG. 8, a part 54 of the gap 54 of the DNA chip 50 is used as an injection port for dropping the sample solution.
a is exposed from the cover plate 55. In (2) of FIG. 8, a minute recess 63 is separately provided on the DNA chip 60 as an injection port, and a gap 64 is formed in the communication path 63a.
I have contacted. In (3) of FIG. 8, a cutout portion 78 is provided on the side edge portion of the cover plate 75 as an inlet.

In the spacer-attached DNA chip of the present invention, the solid phase carrier of the DNA chip can be arbitrarily selected from various materials known as solid phase carrier materials such as glass and silicone. When a signal is detected by an electrochemical method, the solid phase carrier is preferably a conductive substrate composed of electrodes and the like. The surface of the solid phase carrier may be subjected to various known treatments such as coating treatment with polycation and contact treatment with silane coupling agent in order to easily and surely fix the DNA fragment. .

As the DNA fragment probe immobilized on the solid phase carrier, various polynucleotides and oligonucleotides that can be used as DNA fragments for a DNA chip can be used. That is, cDNA (mRNA
, A part of cDNA, EST (200 to 300 bp from the 3'end (bp:
PCR which means a cDNA fragment of about base pairs)
Prepared by the method ("PC
R product ”), and synthetic oligonucleotides. For cDNA, 100 to 1000
It is preferably bp, and if it is oligo DNA, it is 2 to
The 80-mer is preferable. In addition, the DNA fragment is
It may be an artificial nucleic acid obtained by converting a phosphodiester bond of DNA into a peptide bond, that is, a peptide nucleic acid (PNA).

The spotting and immobilization of the above DNA fragment probe on the surface of the solid phase carrier can also be carried out by a conventionally known method. The DNA fragments to be spotted are preferably in the range of 10 ² to 10 ⁵ types / cm ^{2 on} the surface of the solid phase carrier. The amount of the DNA fragment is in the range of 1 to 10 ^-15 mol, and the weight is preferably several ng or less.
It is desirable that the DNA fragments fixed in spots have a substantially circular shape and substantially the same size.
The spotted amount per spot is generally 100 pL to 1 μL
, And preferably in the range of 1 to 100 nL. Although the number of spots is simplified to 10 in the above drawings, it is usually several hundred to several thousand.

When the cover plate has a concave shape, for example, a commercially available gap cover glass (manufactured by Matsunami Glass Industry Co., Ltd.) can be used. Further, when it is flat, a conventionally known cover plate for a microscope can be used. As the cover plate, a glass plate (eg, cover glass) or a plate made of a resin material is preferably used, but the cover plate does not necessarily need to be transparent.

The material of the solid phase carrier that can be used in the DNA chip assembly of the present invention, the DNA fragment, and the method for fixing the same are, for example, Japanese Patent Application Nos. 2000-22180 and 2000-2000 by the present applicant. Each specification such as 166384 can be referred to.

The sample nucleic acid fragments used for analysis by the DNA chip with a cover plate of the present invention include DNA fragments and RNA fragments extracted and treated from various cell or tissue samples detectable by conventional DNA chips. be able to. When the labeling method is used, the sample is a cyanine dye (for example, Cy3, C of Cy Dye ^™ series).
(for example, y5) or a fluorescent substance labeled with RI such as ³² P or ³³ P is used. In the hybridization, a sample solution prepared by dissolving or dispersing these sample nucleic acid fragments in an aqueous medium such as SSC (standard salt-citrate buffer) according to various known methods can be used. In the case of using an electrochemical method, an electrochemically active threading intercalator such as a ferrocene naphthalene diimide derivative may be contained in the sample solution.

Upon hybridization, the sample solution containing the sample nucleic acid fragment can be dropped and uniformly spread on the above-described DNA chip assembly of the present invention as follows. Micro recesses and / or DNA chip assembly
Alternatively, when one or more cutouts are provided, the sample solution is dropped on each of the minute recesses and / or the cutouts with the cover plate placed on the DNA chip. The dropped sample solution is caused by the capillary phenomenon in the space between the DNA chip and the cover plate.
The sample solution spreads smoothly in the hybridization execution area, and the sample solution uniformly and sufficiently spreads over all of the numerous spots on the chip. When a part of the gap on the DNA chip side is exposed from the cover plate, by dropping the sample solution into the exposed gap, the sample solution also spreads smoothly in the hybridization execution region and becomes uniform. And spread to each spot enough.

[0038]

[Experimental Example 1] As a solid-phase carrier, one circular minute recess (aperture diameter: 1000 μm, depth: 50 μm)
Flat glass slides (length: 25 mm, width:
75 mm, thickness: 1.0 mm) was prepared. As the cover plate, a concave cover glass (trade name: gap cover glass, length: 24 mm, width: 50 mm, thickness:
0.35 mm, height of gap: 100 μm, manufactured by Matsunami Glass Industry Co., Ltd.) was prepared. A cover plate was placed on the slide glass to form a structure as shown in FIG. 1 (however, there was no DNA fragment spot).

Separately, a hybridization solution (5
40 pL of a mixed solution of x SSC and 0.5 wt% SDS (sodium dodecyl sulfate) solution), and 1000 p of a dye (bromophenol blue) exhibiting light absorption in the visible region.
An aqueous solution containing pm was prepared.

This aqueous solution was dropped into the minute recesses of the solid phase carrier using a GILSON pipette equipped with a sequencer chip to spread the aqueous solution in the gap between the slide glass and the cover plate. Then MC for this assembly
Using PD (MCPD-2000, manufactured by Otsuka Electronics Co., Ltd.), the position (10) corresponding to the spot portion shown in FIG.
The light absorption spectrum of the dye was measured at each location. The obtained results are summarized in FIG.

[Comparative Experimental Example 1] As an assembly, a flat slide glass (length: 25 mm, width: 75 mm) and a flat microscope cover glass (length: 24 mm, width: 50 m)
m, thickness: 0.35 mm) was prepared. After the aqueous solution containing the dye of Experimental Example 1 was dropped on the slide glass, the cover plate was gently placed to spread the aqueous solution on the slide glass. Then, in the same manner as in Experimental Example 1,
The light absorption spectrum of the dye in this assembly was measured.
The obtained results are summarized in FIG.

FIG. 9 and FIG. 10 are graphs showing the light absorption spectra of dyes at positions corresponding to respective spots.

As is clear from the results shown in FIG. 9, in the assembly having the structure according to the DNA chip assembly of the present invention (Experimental Example 1), almost the same light absorption was observed regardless of the position on the slide glass. The spectrum, that is, the light absorption rate is shown. From this, it can be seen that the aqueous solution containing the dye spreads almost uniformly on the slide glass. On the other hand, as is clear from the results shown in FIG. 10, in the case of the assembly having a structure conforming to the conventional DNA chip and the cover plate (Comparative Experimental Example 1), the light absorption spectrum greatly differs depending on the position on the slide glass, There was a difference in absorption rate. From this, it can be seen that the aqueous solution containing the pigment spreads unevenly and insufficiently on the slide glass.

[0044]

In the DNA chip with a cover plate of the present invention, a spacer is sandwiched between the cover plate and the DNA chip, and when the two are superposed, a minute space is created between them, thereby causing a capillary phenomenon. Thus, the sample solution can be uniformly and sufficiently spread on all the spots on the DNA chip. Also, by providing a minute recess on the DNA chip side as an injection port for dropping the sample solution, or by providing a cutout portion on the cover plate side, the sample solution can be dropped while the cover plate is mounted from the beginning. You can Thereby, it is possible to prevent air bubbles from being mixed when the cover plate is covered later, or to avoid spilling of the sample solution when the sample is simply injected into the gap between the cover plate and the DNA chip. Therefore, hybridization can be uniformly and sufficiently performed in each spot, and the detection accuracy of the DNA chip can be improved. Furthermore, since the cover plate is installed and the position of the sample injection port is fixed in the DNA chip with a cover plate of the present invention, by using this DNA chip assembly, hybridization from dropping of the sample solution to incubation is performed. It is possible to completely automate the operation.

[Brief description of drawings]

FIG. 1 (1) is a DNA with a cover plate of the present invention.
It is a top view which shows an example of a chip | tip, (2) is sectional drawing which follows the II line, and (3) is sectional drawing which follows the II-II line.

2 (1) to (3) are top views each showing an example of variations in the shape of the opening of the minute recess.

3A and 3B are cross-sectional views showing examples of variations in cross-sectional shape of minute recesses.

4 (1) to (5) are top views each showing an example of a variation of a minute recess.

FIG. 5 is a top view showing another example of the DNA chip with a cover plate of the present invention.

FIG. 6 is a top view showing another example of the DNA chip with a cover plate of the present invention.

FIG. 7 (1) is a DNA with a cover plate of the present invention.
FIG. 6 is a top view showing another example of a chip, and (2) is
It is sectional drawing which followed the II line.

8 (1) to (3) are top views each schematically showing another example of the DNA chip with a cover plate of the present invention.

FIG. 9 is a graph showing a light absorption spectrum of a dye at positions corresponding to respective spots in an analytical tool having a configuration according to the present invention.

FIG. 10 is a graph showing a light absorption spectrum of a dye at positions corresponding to respective spots in an analytical tool having a configuration according to the related art.

[Explanation of symbols]

10, 20, 30, 40, 50, 60, 70 DNA chip 11, 21, 31, 41 Solid phase carrier 12, 22, 32, 42 Spot 13, 33, 63 Micro recess 44, 54, 64, 74 Void 15, 25, 35, 45, 55, 65, 75 Cover plate 16, 17, 26, 27, 36, 37 End portion 28, 38, 78 Cutout portion 16, 17, 41a Spacer

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Osamu Soshimoto             3-11-46 Izumi, Asaka-shi, Saitama Fuji Photographic Group             Within Ilum Co., Ltd. F-term (reference) 4B024 AA11 HA11 HA12 HA19                 4B029 AA07 BB01 CC03 DC07 FA02                       FA15 GA08 GB05 GB06 GB09                       HA05

Claims (21)

[Claims] 1. A D having an upper surface having a hybridization execution region including a plurality of DNA fragment probe fixing spots and a sample solution spotting region arranged in contact with the region.
DNA with a cover plate, which comprises an NA chip and a cover plate provided above the hybridization execution region of the DNA chip with an interval that allows the aqueous DNA fragment sample solution to spread to the hybridization region by capillary action In the chip, the sample solution spotting hole portion is provided on the boundary between the sample solution spotting area and the hybridization execution area, and at a position corresponding to the lower side of one side of the cover plate adjacent to the boundary. Characteristic DNA chip with cover plate. 2. The DNA chip with a cover plate according to claim 1, wherein the cover plate is made of a transparent glass material or a resin material. 3. The DNA chip with a cover plate according to claim 1, wherein the distance between the DNA chip and the cover plate is in the range of 5 to 200 μm. 4. The cover plate according to claim 1, wherein the diameter of the opening of the hole for spotting the sample liquid of the DNA chip is in the range of 100 to 2000 μm, and the depth of the hole is in the range of 5 to 200 μm. With DNA chip. 5. The DNA chip with a cover plate according to claim 1, wherein the sample solution spotting hole is a groove-like hole formed along the side of the cover plate. 6. A DNA chip having an upper surface with a hybridization execution area and a sample solution spotting area arranged in contact with the area, a cover plate, and a capillary phenomenon above the hybridization execution area of the DNA chip. A spacer for arranging a cover plate with an interval that allows the aqueous DNA fragment sample solution to spread to the hybridization region, and the sample is provided on the boundary between the sample solution spotting region and the hybridization execution region. D characterized by having a liquid spot wearing hole
NA chip assembly. 7. The DNA chip according to claim 6, wherein the diameter of the opening of the hole for spotting the sample solution of the DNA chip is in the range of 100 to 2000 μm, and the depth of the hole is in the range of 5 to 200 μm. Assembly. 8. The DNA chip assembly according to claim 6, wherein the sample solution spotting hole is a groove-shaped hole formed along the boundary. 9. The spacer is a cover plate or DN.
The DNA chip assembly according to claim 6, which is fixed to both side portions of the A chip. 10. A DNA chip having, on its upper surface, a hybridization execution area and a sample solution spotting area arranged in contact with the area, on the boundary between the sample solution spotting area and the hybridization execution area. A DNA chip having a hole for spotting a sample liquid. 11. The diameter of the opening of the sample liquid spot wearing hole is 1
The DNA according to claim 10, wherein the DNA is in the range of 00 to 2000 µm and the depth of the hole is in the range of 5 to 200 µm.
Chips. 12. The DNA chip according to claim 10, wherein the sample solution spotting hole is a groove-like hole formed along the boundary. 13. A DNA chip having an hybridization surface including a plurality of DNA fragment probe fixing spots and a sample solution spotting area arranged in contact with the area, and a hybridization of the DNA chip. Aqueous DNA by capillary action above the working area
In a DNA chip with a cover plate, which comprises a cover plate provided with an interval that allows the fragment sample solution to spread to the hybridization region, above the boundary between the sample solution spotting region and the hybridization execution region. A DNA chip with a cover plate, characterized in that one side of the cover plate located is provided with an opening or a notch for spotting the sample liquid. 14. The DNA chip with a cover plate according to claim 13, wherein the cover plate is made of a transparent glass material or a resin material. 15. The DNA chip with a cover plate according to claim 13, wherein the distance between the DNA chip and the cover plate is in the range of 5 to 200 μm. 16. The DN with cover plate according to claim 13, wherein the diameter of the opening or notch for wearing the sample liquid spot on the cover plate is in the range of 100 to 2000 μm.
A chip. 17. The sample solution spotting hole is also provided on the surface of the DNA chip located below the opening or notch for the sample solution spotting of the cover plate.
A DNA chip with a cover plate according to 1. 18. A DNA chip, a cover plate, and a DNA having an upper surface having a hybridization execution region and a sample solution spotting region arranged in contact with the region.
A spacer is provided above the hybridization execution region of the chip so as to arrange the cover plate with an interval that allows the aqueous DNA fragment sample solution to spread to the hybridization region by capillary action, and the sample is provided on one side of the cover plate. A DNA chip assembly comprising an opening or notch for wearing a liquid spot. 19. The DNA chip assembly according to claim 18, wherein the diameter of the opening or notch for wearing the sample liquid spot on the cover plate is in the range of 100 to 2000 μm. 20. The spacer is a cover plate or D
The DNA chip assembly according to claim 18, which is fixed to both side edges of the NA chip. 21. A cover plate having spacers on both sides, and an opening or a notch having a diameter in the range of 100 to 2000 μm is formed on a side other than the side.