JP2002181817A - Method for producing polynucleotide array - Google Patents

Abstract

(57) [Summary] (Corrected) [PROBLEMS] To prevent cross-contamination of various kinds of polynucleotides spotted on a support while firmly fixing the concentration of the polynucleotides to be substantially uniform in a substantially uniform state. Provided is a method for producing a polynucleotide array having sufficient measurement sensitivity by performing an analysis operation effectively and efficiently. [MEANS FOR SOLVING PROBLEMS] 1. Prepare a solution by dissolving the polynucleotide in at least about 10% by volume of formamide in water. The above solution was used in (1). Wash the tip of the dispensing needle with the washing liquid, (2). Sampling the solution on a dispensing needle, (3). (4) The dispensing needle is brought into contact with the discard plate to remove excess solution adhering to the tip. A large number of the solutions are spotted by abutting a dispensing needle on the surface of the support (5). Drying to fix the polynucleotide, (6).
The support is immersed in a blocking solution to block the surface of the support between the polynucleotide spots. The polynucleotide is immobilized on the surface of the support according to the above-described steps.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a polynucleotide array in which many and many kinds of polynucleotides are immobilized in spots on a support.

[0002]

In the study of gene expression and gene polymorphism analysis, a suitable salt (0.45 mol / l sodium chloride,
Polynucleotide arrays (DNA) in which a large number of various types of polynucleotide solutions obtained by dissolving different types of polynucleotides in respective solutions containing 0.045 mol / l sodium citrate, etc., are spotted at high density and dried and fixed. Chip), so that multiple types of polynucleotide samples can be analyzed simultaneously on multiple items in one measurement operation.

[0003] However, in the above-mentioned polynucleotide array, after the dried and fixed polynucleotide is dried and fixed, a blocking treatment for preventing the test nucleotide from adhering to the surface of the support between the polynucleotides is performed. In addition, when the nucleotide solution for test was dropped on the surface of the support for hybridization, the polynucleotide was easily peeled off from the surface of the support, and the nucleotide for test could not be measured effectively and with high sensitivity.

[0004] This disadvantage can be solved by treating the surface of the support so as to easily bind to amino groups, for example, and aminating the end of the polynucleotide to be fixed to increase the fixing strength. Amination and modification of the termini of the polynucleotides require a great deal of cost and time, resulting in an increase in analysis costs.

[0005] Further, in order to increase the reaction time between the surface of the support and the polynucleotide to enhance the immobilization reaction, a treatment using a highly water-absorbing solvent such as dimethyl sulfoxide or ethylene glycol as the solvent in which the polynucleotide is dissolved. A method has been proposed.

In this method, when a polynucleotide is spotted on a support, the spot shape is likely to be non-uniform, and the concentration of the polynucleotide is non-uniform, so that the test nucleotide can be obtained with high sensitivity. It was difficult to analyze efficiently.

Further, as a device for spotting a large number of polynucleotide solutions in a predetermined amount on the surface of a support, a dispensing needle provided with a longitudinally extending groove at the tip end thereof is used. A predetermined amount of the polynucleotide is spotted in a predetermined spot shape by contacting the surface of the support after sampling the polynucleotide solution.

However, in the spotting method using the dispensing needle described above, at the initial stage of spotting, the excess polynucleotide solution adhering to the tip portion flows out and adheres to the support, and the spot diameter increases. Therefore, there is a problem that the spots of the polynucleotide may be contaminated with each other, and the concentration of the spots is not made uniform due to the non-uniform spot shape, so that the nucleotide to be tested cannot be detected with high sensitivity.

Further, in the spotting method using the above-described dispensing needle, a single dispensing needle spots many kinds of polynucleotide solutions, but the dispensing needle is not sufficiently washed. In the above, the polynucleotide to be spotted was contaminated and the test nucleotide could not be accurately analyzed.

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned conventional drawbacks, and an object of the present invention is to prevent various kinds of polynucleotides spotted on a support while preventing cross-contamination. It is an object of the present invention to provide a method for producing a polynucleotide array that can effectively and efficiently perform a work of analyzing a test nucleotide by firmly fixing a polynucleotide in a substantially uniform concentration state.

Another object of the present invention is to provide a method for producing a polynucleotide array capable of obtaining a sufficient measurement sensitivity by making the spot concentration of a polynucleotide sample spotted on a support substantially uniform. It is in.

[0012]

According to the first aspect of the present invention, there is provided a method for producing a polynucleotide array by abutting a dispensing needle on a surface of a support, spotting a polynucleotide solution, and then fixing the solution. 1. preparing a polynucleotide solution by dissolving the polynucleotide in at least about 10% by volume of formamide in water. The polynucleotide prepared as described above is characterized in that the polynucleotide is immobilized on the surface of a support according to the following steps.

(1). The tip of the dispensing needle is washed with the washing liquid. (2). The polynucleotide solution is sampled on the dispensing needle whose tip has been washed. (3). The dispensing needle from which the polynucleotide solution has been sampled is discarded, and the surplus polynucleotide solution adhering to the tip is removed by contacting the hitting plate. (Four). A number of polynucleotide solutions are spotted by contacting the dispensing needle against the surface of the support. (Five). A large number of polynucleotide solutions spotted on the surface of the support are dried to fix the polynucleotides. (6). The support on which the polynucleotide is immobilized is immersed in a blocking solution to block the surface of the support between the polynucleotide spots.

According to a second aspect of the present invention, there is provided a method for producing a polynucleotide array by abutting a dispensing needle on the surface of a support, spotting a polynucleotide solution, and then fixing the solution. Dissolving the polynucleotide in a solvent in which at least about 10% by volume of glycerol is added to water to prepare a polynucleotide solution; immobilizing the polynucleotide prepared as described above on the surface of a support according to the following steps: It is characterized by.

(1). The tip of the dispensing needle is washed with the washing liquid. (2). The polynucleotide solution is sampled on the dispensing needle whose tip has been washed. (3). The dispensing needle from which the polynucleotide solution has been sampled is discarded, and the surplus polynucleotide solution adhering to the tip is removed by contacting the hitting plate. (Four). A number of polynucleotide solutions are spotted by contacting the dispensing needle against the surface of the support. (Five). A large number of polynucleotide solutions spotted on the surface of the support are dried to fix the polynucleotides. (6). The support on which the polynucleotide is immobilized is immersed in a blocking solution to block the surface of the support between the polynucleotide spots.

[0016]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be described below with reference to the drawings. 1 and 2, the support 3 of the polynucleotide array 1 can be made of, for example, glass, membrane, plastic (polyethylene, polypropylene, polystyrene, etc.), and a preferred example is manufactured by Matsunami Glass Co., Ltd. A slide glass (model S9115) can be used.

Surface Treatment of Support The surface of the support 3 is subjected to a positive charge treatment or an amination treatment. As the positive charge treatment, the surface of the support 3 is surface-treated with poly-L-lysine, aminopropyltriethoxysilane, or the like to be positively charged. In the amination treatment, the surface of the support 3 which has been positively charged by the above method is treated with an aldehyde or succinide so that the amino group is easily bonded. This amination treatment is effective in promoting the binding of the amino group at the terminal of the polynucleotide to be immobilized.

Preparation of Polynucleotide Solution A polynucleotide solution to be fixed on the surface of the support 3 is prepared as follows.

The kind of the polynucleotide (nucleic acid) used in the present invention is not particularly limited, and includes DNA (deoxyribonucleic acid), RNA (ribonucleic acid) and the like. And DN
A and RNA are not particularly limited, and chromosomal DNA
DNA fragments, mRNA, cDNA, chemically synthesized DN
A or any of chemically synthesized RNAs.
Any of a CR amplification product, restriction enzyme digested DNA and the like may be used.

The polynucleotide used in the present invention may be derived from any organism. The polynucleotide used in the present invention includes a DNA fragment derived from a chromosome position known to be associated with a disease, a cDNA fragment, and a chemically synthesized DNA. Furthermore, the polynucleotide of the present invention may be a DNA fragment containing a part of a specific gene, mRNA having an abnormal transcription level, or cDNA derived from the mRNA.

The above-mentioned polynucleotide of the present invention can be prepared using pure water or a 3 × SSC solution (0.45 M sodium chloride,
10% by volume of formamide or glycerol was added to the mixture to adjust the polynucleotide concentration to 0.5 to 50 pmol / μl.
Use the one adjusted to.

In order to spot the polynucleotide solution on the surface of the support 3, a large number of dispensing needles 5
However, dispensing heads 7 arranged in a matrix at intervals of, for example, 100 to 200 μm are used. A large number of polynucleotide solutions are spotted by controlling the movement in the XYZ axis direction with respect to the surface of the support 3. Each dispensing needle 5 used for this spotting has a tip diameter of, for example, 100 to 20.
At 0 μm, a vertically dividing groove 5a is formed in the axial direction from the tip, and the polynucleotide solution spotted in the vertically dividing groove 5a is absorbed by capillary action and stored.

The dispensing needle 5 has a through-hole at the tip and a heating member attached to the outer periphery of the tip, and a polynucleotide solution stored in a tank provided at the rear end and retained in the through-hole. May be spotted by instantaneously heating the heating member to form bubbles and pop out on the surface of the support 3.

Further, in addition to the above-described dispensing needle of the bubble pop-out type, a type in which the polynucleotide solution is directly popped out by using deformation of a piezo element or ultrasonic vibration and spotted on the surface of the support 3 may be used. Good.

When the polynucleotide solution is spotted on the surface of the support 3 by controlling the movement of the dispensing head 7, first, the tip of each dispensing needle 5 in the dispensing head 7 is washed and spotted. Prevents contamination of various types of polynucleotides (carryover).

As the cleaning method, first, the tip of each dispensing needle 5 in the dispensing head 7 is pressed against a cleaning water-absorbing plate such as a porous ceramic plate or a water-absorbing paper plate to remove the poly remaining in each vertically-divided groove 5a. After discharging the nucleotide solution, the solution is immersed in the order of a 0.1% sodium dodecyl sulfate aqueous solution tank, an ultrapure water tank, and an ethanol tank for about 1 to 5 sec. This is performed by drying the attached ethanol by phase suction.

Sampling of Polynucleotides The tip of each dispensing needle 5 in the dispensing head 7 that has been washed as described above is filled with various kinds of polynucleotide solutions to be fixed to the surface of the support 3. The polynucleotide solution is immersed in each solution tank of the 384-well multi-plate for about 7 sec., And the polynucleotide solution is absorbed into each of the vertically divided grooves 5a by capillary action and sampled.

Removal of surplus polynucleotide Prior to the spotting operation by the dispensing head 7, a discard operation is performed. That is, when the polynucleotide solution is sampled on the dispensing needle 5, the surplus polynucleotide solution remains attached to the outer surface of the tip of each dispensing needle 5. For this reason, the tip surface of each dispensing needle 5 is pressed several times at a time of about 0.5 to 1 sec. Each time against a discarded punching plate such as a porous ceramic plate, a water-absorbing paper plate or a glass plate. Excess solution adhering to the outer surface of the tip of the dispensing needle 5 is absorbed.

This prevents excess polynucleotide solution adhering to the tip of the dispensing needle 5 from being spotted at the beginning of spotting on the surface of the support 3 and reducing the size of each spot due to the polynucleotide solution to be spotted. Substantially uniformity makes the concentration of the polynucleotide solution uniform and prevents contamination between spots.

Next, the dispensing head 7 is moved to the surface of the support 3 by XYZ
Each dispensing needle 5 in the dispensing head 7 is controlled to move in the axial direction.
The various types of polynucleotide solutions are spotted in a matrix by contacting the tip surfaces of the above for about 0.5 to 1 sec. In this spotting operation, the dispensing head 7
-The distance between the spots is approximately 100 to
The movement is controlled to be 200 μm. The spotting environment at that time may be a normal temperature environment, but preferably the external environment temperature is about 2 to 10 ° C, preferably about 2 to 6 ° C, and the humidity is kept at 50 to 70%. By doing so, the spotted polynucleotide solution can be prevented from drying in a short time, and the immobilization time can be lengthened.

More specifically, the entire area including the support 3 and the dispensing head 7 is partitioned from the outside air, and a temperature sensor and a humidity sensor are provided in the partitioned internal area so that the heating and heating are performed so that the above-mentioned set temperature and humidity are obtained. This is performed by providing a humidifying device.

Next, water vapor is applied to the surface of the support 3 on which a large number of polynucleotide solutions of various types are spotted.
After spraying for 3 seconds, the support 3 is placed on a hot plate heated to about 80 to 95 ° C. and dried and fixed. When the tip surface of the dispensing needle 5 is brought into contact with the surface of the support 3 and the polynucleotide solution is spotted, the adhesion state of the polynucleotide solution in each spot becomes uneven, and the concentration in the spot is not constant. In order to avoid this, after spotting the polynucleotide solution on the support 3, the concentration of the spotted polynucleotide solution is reduced by spraying water vapor to make the adhesion state at each spot uniform.

At this time, in the polynucleotide solution to which formamide is added, a certain form can be maintained by inhibiting the denaturation of nucleotides by formamide. Therefore, the positively charged support 3
When spotting is performed on the surface of the substrate 3, the probability that the negatively charged phosphate in the polynucleotide is electrostatically bonded to the surface of the support 3 is increased.

Further, when spotting is performed on the surface of the aminated support 3, the probability of binding to the amino group at the end of the polynucleotide is increased. That is, when the polynucleotide is constantly denatured, it becomes difficult to form an electrostatic bond or an amino group bond. However, the denaturation of the polynucleotide is inhibited and the binding probability can be increased by keeping the polynucleotide in a certain form.

Furthermore, in the polynucleotide solution to which glycerose is added, the denaturation of nucleotides is not inhibited as in the case of formamide addition, and is constantly denatured. By increasing the time, the connection probability can be increased.

Thus, the fixing strength of the polynucleotide to the surface of the support 3 is increased.

Drying Next, a UV crosslinker (UVC5 manufactured by Hoefer) was applied to the surface of the support 3 on which the polynucleotide was immobilized.
(00) to irradiate about 50 to 120 mJ of ultraviolet light to immobilize the polynucleotide.

Next, the non-spotting region of the polynucleotide on the surface of the support 3 is subjected to a blocking treatment. As the blocking treatment method, 1 to 1.5 wt% succinic anhydride, 9
The entire support 3 on which the polynucleotide was immobilized in a block solution adjusted to pH 5 to 6 with 0 vol% N-methylpyrosinone and 10 vol% 0.2 M sodium borate solution (pH: 8.0) for about 15 minutes. During this time, it is immersed.

Then, the support 3 taken out from the block solution is immersed in hot water of 95 to 99 ° C. and left for about 60 seconds, and then the support 3 is immersed in ethanol.
Allow the surface to air dry.

As another blocking treatment method, a blocking solution adjusted to pH 8 to 9 with an ethanolamine solution is used and immersed similarly. That is, the support 3 on which the polynucleotide was spotted was heated at 45 to 65 ° C.
After incubating for 4-14 hours under saturated steam of ethanol, each final concentration 50 mmol / l ethanolamine,
0.1M Tris buffer pH: 9.0, immersed in a block solution adjusted to pH: 8 to 9 with 0.1% sodium dodecyl sulfate for 10 to 30 min.
C. at 15 ° C. for 15 to 60 minutes in an SSC solution containing 0.1% sodium dodecyl sulfate. Furthermore, after washing with water, drying is performed after removing the liquid component adhering to the support 3.

Comparative Example 1 In this comparative example, a synthetic oligonucleotide (80 bases, final concentration 25
pmol / μl) in 3 × SSC solution, phosphate buffer (pH:
7.5) Using a synthetic oligonucleotide solution dissolved in four kinds of solvents of ultrapure water and a 10% by volume aqueous formaldehyde solution, the immobilization rate of each solvent was verified.

The nucleotides used for the hybridization were oligonucleotides (80 bases, final concentration of 1) having a complementary nucleotide sequence to the synthetic oligonucleotide fixed on the surface of the support 3 and labeled with a fluorescent dye (Cy3).
pmol / μl, containing 0.1 g / ml salmon sperm DNA).

As the support 3, a slide glass for DNA chips manufactured by Matsunami Glass Industry Co., Ltd. (trade name: M
AS-coated slide).

As shown in FIGS. 3 and 4, when the synthetic oligonucleotide is dissolved in formaldehyde as a solvent, the fluorescence intensity is highest as compared with other solvents. This fact indicates that when formaldehyde is used as a solvent for the synthetic oligonucleotide, the amount of the synthetic oligonucleotide immobilized on the surface of the support 3 is the largest.

FIG. 5 shows a support obtained when a blocking step and a hybridization step were performed on a polynucleotide array 1 in which a synthetic oligonucleotide solution obtained by dissolving a synthetic oligonucleotide using formamide as a solvent was spotted and fixed. This is the result of 16 trials of the immobilization rate of the synthetic oligonucleotide spot on the 3 surfaces. When formamide is used as a solvent, the average immobilization rate is 97.6%.
Thus, a sufficient fixed amount could be left.

Comparative Example 2 FIGS. 6 and 7 show a comparison between the case where discarding is performed prior to spotting and the case where no dumping is performed. In this experiment, a porous ceramic plate was used as the water absorbing plate 9 and the support 3 was used. A λ phage DNA labeled with a fluorescent dye (Cy3) was used as a polynucleotide to be spotted on the surface of the sample.

Then, after 116 spots were continuously spotted on the surface of the support 3 by one spotting operation, the fluorescence intensity was read by a scanner and graphed.

As a result, in the case where tapping is performed several times before spotting on the surface of the support 3, the fluorescence intensity per spot is made uniform as compared with the case where tapping is not performed. Conversely, no tapping indicates that the fluorescence intensity at the initial stage of spotting is high, and therefore the amount of polynucleotide spots is large.

Comparative Example 3 Using one dispensing needle 5, a fluorescent solution in which 400 fmol / μl of a fluorescent dye (Cy3, Cy5) was dissolved in pure water and pure water were alternately sampled, and the surface of the support 3 was sampled. And fluorescent labels are detected by a scanner. The spotting cycle is fluorescence sampling → spotting →
Washing, pure water sampling, spotting, etc. are performed in this order, and the washing effect of the dispensing needle 5 is verified.

As shown in FIG. 8, since spots where fluorescence is detected and spots where no fluorescence are detected appear alternately, the inside of the vertical groove 5a is almost completely washed by washing the dispensing needle 5 described above. This indicates that no fluorescent dye was detected in pure water.

[0051]

Industrial Applicability The present invention provides an analysis of nucleotides to be tested by firmly fixing various types of polynucleotides in a substantially uniform concentration while preventing cross-contamination of polynucleotides spotted on a support. Work can be performed effectively and efficiently. Further, the spot concentration of the polynucleotide sample spotted on the support can be made substantially uniform to obtain sufficient measurement sensitivity.

[Brief description of the drawings]

FIG. 1 is an explanatory diagram showing a process for producing a polynucleotide array.

FIG. 2 is a partial perspective view showing a spotting state of the polynucleotide array.

FIG. 3 is a photograph obtained by observing, with a scanner, a fluorescence state of each solvent when hybridization is performed using a polynucleotide dissolved in each solvent.

FIG. 4 is a graph quantifying the fluorescence intensity of each spot in FIG.

FIG. 5 is a table showing the immobilization ratio of a polynucleotide according to the number of times of a blocking process and a hybridization process.

FIG. 6 is a graph showing the fluorescence intensity when a dispensing needle is tapped before spotting a polynucleotide on a support.

FIG. 7 is a graph showing the fluorescence intensity when spotting is performed without tapping.

FIG. 8 is a photograph showing a cleaning effect of a dispensing needle.

[Explanation of symbols]

1-polynucleotide array, 3-support, 5-dispensing needle, 5a-vertical groove

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[Procedure amendment]

[Submission date] July 18, 2001 (2001.7.1)
8)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0010

[Correction method] Change

[Correction contents]

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned conventional drawbacks, and an object of the present invention is to prevent various kinds of polynucleotides spotted on a support while preventing cross-contamination. It is an object of the present invention to provide a method for producing a polynucleotide array that can effectively and efficiently perform a work of analyzing a test nucleotide by firmly fixing a polynucleotide in a substantially uniform concentration state.

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0029

[Correction method] Change

[Correction contents]

This prevents excess polynucleotide solution adhering to the tip of the dispensing needle 5 from being spotted at the beginning of spotting on the surface of the support 3 and reducing the size of each spot due to the polynucleotide solution to be spotted. It is made substantially uniform to make the concentration of the polynucleotide solution uniform and to prevent contamination between spots.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 37/00 102 C12N 15/00 F

Claims (16)

[Claims] 1. A method for producing a polynucleotide array, comprising the steps of: (1) contacting a dispensing needle to the surface of a support, spotting a polynucleotide solution, and fixing the polynucleotide solution to produce a polynucleotide array; 1. The polynucleotide is dissolved in a solvent containing at least about 10% by volume formamide in water to prepare a polynucleotide solution. 2. The polynucleotide is immobilized on the surface of the support according to the following steps. (1). The tip of the dispensing needle is washed with the washing liquid. (2). The polynucleotide solution is sampled on the dispensing needle whose tip has been washed. (3). The dispensing needle from which the polynucleotide solution has been sampled is discarded and abuts against the striking plate to remove excess polynucleotide solution attached to the tip. (Four). A number of polynucleotide solutions are spotted by contacting the dispensing needle against the surface of the support. (Five). A large number of polynucleotide solutions spotted on the surface of the support are dried to fix the polynucleotides. (6). The support on which the polynucleotide is immobilized is immersed in a blocking solution to block the surface of the support between the polynucleotide spots. 2. A method for producing a polynucleotide array, comprising the steps of: contacting a dispensing needle to the surface of a support, spotting a polynucleotide solution, and fixing the solution; 1. The polynucleotide is dissolved in a solvent containing at least about 10% by volume of glycerol in water to prepare a polynucleotide solution. 2. The polynucleotide is immobilized on the surface of the support according to the following steps. (1). The tip of the dispensing needle is washed with the washing liquid. (2). The polynucleotide solution is sampled on the dispensing needle whose tip has been washed. (3). The dispensing needle from which the polynucleotide solution has been sampled is discarded and abuts against the striking plate to remove excess polynucleotide solution attached to the tip. (Four). A number of polynucleotide solutions are spotted by contacting the dispensing needle against the surface of the support. (Five). A large number of polynucleotide solutions spotted on the surface of the support are dried to fix the polynucleotides. (6). The support on which the polynucleotide is immobilized is immersed in a blocking solution to block the surface of the support between the polynucleotide spots. 3. The method for producing a polynucleotide array according to claim 1, wherein the concentration of the polynucleotide dissolved in the solvent is 0.5 to 50 pmol / μl. 4. The method for producing a polynucleotide array according to claim 1, wherein the polynucleotide is a PCR amplification product derived from a biological gene. 5. The method according to claim 1, wherein the polynucleotide is an artificially synthesized oligonucleotide. 6. The method for producing a polynucleotide array according to claim 1, wherein the discarded plate is any one of a porous ceramics plate, a water-absorbing paper plate and a glass plate. 7. The washing of the dispensing needle according to claim 1 or 2, wherein the dispensing needle is immersed in a 0.1% sodium dodecyl sulfate solution for discharging the polynucleotide solution remaining in the vertically divided groove by contacting with a water absorbing material. A method for producing a polynucleotide array, comprising: immersing in pure water, immersing in ethanol, and vapor-phase suction-drying. 8. The method for producing a polynucleotide array according to claim 1, wherein the environment for spotting the polynucleotide on the surface of the support is a normal temperature environment. 9. The method for producing a polynucleotide array according to claim 1, wherein the environment for spotting the polynucleotide on the surface of the support is 2 to 10 ° C. and 50 to 70% humidity. 10. The method according to claim 1, wherein the surface of the support is positively charged by surface treatment with either poly-L-lysine or aminopropyltriethoxysilane. 11. The method according to claim 1, wherein the surface of the support is aminated by either aldehyde treatment or succinide treatment. 12. The method according to claim 1, wherein the blocking solution is a succinic acid solution adjusted to at least pH 5 to 6. 13. The method for producing a polynucleotide array according to claim 1, wherein the blocking solution is an ethanolamine solution adjusted to at least pH 8-9. 14. The method for producing a polynucleotide array according to claim 1, wherein water vapor is sprayed onto the surface of the support on which the polynucleotide solution has been spotted to make the adhesion state of the polynucleotide solution uniform at each spot. 15. A method for producing a polynucleotide array according to claim 1 or 2, wherein the dispensing needle has a longitudinally extending groove formed at an end thereof extending in the axial direction, and a polynucleotide solution can be sampled in the longitudinally dividing groove. . 16. A dispensing needle according to claim 1, wherein the dispensing needle is provided with a heating member at a tip portion having a through-hole, and the polynucleotide solution in the through-hole is instantaneously heated by the heating member to form bubbles to form a support. A method for producing a polynucleotide array in which a polynucleotide solution is spotted on a surface.