Inkjet method of spotting probe and manufacturing a probe array

• US 20030059817A1
• Filed: 08/16/2002
• Published: 03/27/2003
• Est. Priority Date: 08/01/1997
• Status: Abandoned Application

• Current Assignee: Nobuko Yamamoto, Tadashi Okamoto, Tomohiro Suzuki
• Original Assignee: Nobuko Yamamoto, Tadashi Okamoto, Tomohiro Suzuki
• Inventors: Suzuki, Tomohiro, Okamoto, Tadashi, Yamamoto, Nobuko
• Application Number:
  US10/219,313
• Publication Number:
  US 20030059817A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00274 Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 2219/00729 : Peptide nucleic acids [PNA]
• CPC Class Codes: B01L 2200/143 : Quality control, feedback systems
• CPC Class Codes: B01L 2400/0442 : thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 3/0268 : using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 : Compounds covalently bound to a solid support
• CPC Class Codes: C07H 21/00 : Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12N 11/14 : Enzymes or microbial cells immobilised on or in an inorganic carrier
• CPC Class Codes: C12N 15/11 : DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• CPC Class Codes: C12N 2310/351 : Conjugate
• CPC Class Codes: C12Q 1/00 : Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C40B 30/04 : by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/68 : involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/6845 : Methods of identifying protein-protein interactions in protein mixtures

Abstract

Provided is a method of spotting a probe densely and efficiently on a surface of a solid support. A liquid containing a probe is attached to a solid support as droplets to form spots containing the probe on the solid support by an ink jet method.

Claims

1. A method of spotting a probe which can bind specifically to a target to a solid support comprising the steps of:

supplying a liquid containing a probe on a surface of a solid support by an ink jet method and attaching the liquid, and

forming a spot of probe on the surface of the solid support.

2. The method of spotting according to claim 1 wherein the probe is a single-stranded nucleic acid probe.

3. The method of spotting according to claim 2 wherein the single-stranded nucleic acid probe includes a single-stranded DNA probe.

4. The method of spotting according to claim 2 wherein the single-stranded nucleic acid probe includes an RNA probe.

5. The method of spotting according to claim 2 wherein the single-stranded nucleic acid probe includes a single-stranded PNA probe.

6. The method of spotting according to claim 2 wherein the surface of the solid support has a first functional group and the single-stranded nucleic acid probe has a second functional group, and the first and second functional groups react each other by contact.

7. The method of spotting according to claim 6 wherein the first functional group on the surface of the solid support is a maleimido group and the second functional group of the single-stranded nucleic acid probe is a thiol (SH) group.

8. The method of spotting according to claim 7 wherein the solid support is a glass plate and the maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with N-(6-maleimidocaproyloxy) succinimide.

9. The method of spotting according to claim 7 wherein the solid support is a glass plate and the maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with succinimidyl-4-(maleimido phenyl) butyrate.

10. The method of spotting according to claim 7 wherein the maleimido group is reacted with the thiol group for at least 30 minutes.

11. The method of spotting according to claim 10 wherein the single-stranded nucleic acid comprises a single-stranded PNA probe, at the terminus of which the thiol group exists, and the maleimido group is reacted with the thiol group for at least 2 hours.

12. The method of spotting according to claim 11 wherein the thiol group at the terminus of the single-stranded PNA probe is introduced by binding cysteine group to the N-terminus of the single-stranded PNA probe.

13. The method of spotting according to claim 6 wherein the first functional group on the surface of the solid support is an epoxy group and the second functional group of the single-stranded nucleic acid probe is an amino group.

14. The method of spotting according to claim 13 wherein the solid support is a glass plate and the epoxy group is introduced by applying a silane compound having an epoxy group in the molecule thereof on the surface of the glass plate and reacting the compound with the glass plate.

15. The method of spotting according to claim 13 wherein the epoxy group is introduced by applying polyglycidyl methacrylate having an epoxy group on the solid support.

16. The method of spotting according to claim 1 wherein the liquid contains urea at 5-10 wt %, glycerin at 5-10 wt %, thiodiglycol at 5-10 wt %, and an acetylene alcohol at 1 wt % of the liquid.

17. The method of spotting according to claim 16 wherein the acetylene alcohol has a structure represented by the following general formula (I):

18. The method of spotting according to claim 2 wherein a concentration of the single-stranded nucleic acid probe in the liquid is 0.05-500 μ

M.

19. The method of spotting according to claim 18 wherein a concentration of the single-stranded nucleic acid probe in the liquid is 2-50 μ

M.

20. The method of spotting according to claim 2 wherein a length of the single-stranded nucleic acid probe is 2-5,000 bases.

21. The method of spotting according to claim 20 wherein a length of the single-stranded nucleic acid probe is 2-60 bases.

22. The method of spotting according to claim 1 wherein the ink jet method is a bubble jet method.

23. The method of spotting according to claim 1 wherein the probe is an oligopeptide or a polypeptide with a specific amino acid sequence.

24. The method of spotting according to claim 1 wherein the probe is a protein.

25. The method of spotting according to claim 24 wherein the protein is an antibody.

26. The method of spotting according to claim 24 wherein the protein is an enzyme.

27. The method of spotting according to claim 1 wherein the probe is an enzyme.

28. The method of spotting according to claim 1 wherein the liquid is supplied so as to form independent spots in a density of 10,000 spots per square inch on the solid support.

29. The method of spotting according to claim 1 wherein the solid support has a flat surface and homogenous surface properties.

30. The method of spotting according to claim 29 wherein the liquid is supplied on the surface of the solid support so as to obtain a distance between the adjacent spots not smaller than the maximum width of the spot.

31. The method of spotting according to claim 30 wherein blocking is performed on the surface of the solid support to prevent a sample from attaching to the surface other than spots of the surface of the solid support.

32. The method of spotting according to claim 31 wherein the blocking is achieved by using bovine serum albumin.

33. The method of spotting according to claim 1 wherein the solid support is partitioned by a matrix arranged in a pattern on the surface, a plurality of wells whose bottom is the surface of the solid support exposed in the pattern are provided, and the liquid is supplied to the respective wells.

34. The method of spotting according to claim 33 wherein the solid support is optically transparent and the matrix is opaque.

35. The method of spotting according to claim 33 wherein the matrix comprises a resin.

36. The method of spotting according to claim 33 wherein the surface of the matrix is hydrophobic.

37. The method of spotting according to claim 33 wherein the bottom of the wells is hydrophilic.

38. The method of spotting according to claim 33 wherein the matrix has a thickness of 1-20 μ

m.

39. The method of spotting according to claim 33 wherein the wells have a maximum width of 200 μ

m.

40. The method of spotting according to claim 33 wherein the matrix has a width ½

-2 times the maximum width of the wells.

41. A probe array comprising a plurality of spots of a probe, the spots being provided independently at a plurality of sites of a surface of a solid support in a density of 10,000 spots per square inch or higher.

42. The probe array according to claim 41 wherein the solid support has a flat surface and homogenous surface properties.

43. The probe array according to claim 42 wherein the probe is a single-stranded nucleic acid probe.

44. The probe array according to claim 43 wherein the single-stranded nucleic acid probe includes a single-stranded DNA probe.

45. The probe array according to claim 43 wherein the single-stranded nucleic acid includes a single-stranded RNA probe.

46. The probe array according to claim 43 wherein the single-stranded nucleic acid includes a single-stranded PNA probe.

47. The probe array according to claim 43 wherein the single-stranded nucleic acid is covalently bound to the surface of the solid support by a reaction between a first functional group on the surface of the solid surface and a second functional group of the single-stranded nucleic acid probe.

48. The probe array according to claim 47 wherein the first functional group on the surface of the solid support is a maleimido group and the second functional group of the single-stranded nucleic acid probe is a thiol (SH) group.

49. The probe array according to claim 48 wherein the single-stranded nucleic acid probe is a single-stranded PNA probe and contains a cysteine residue on an N-terminus side.

50. The probe array according to claim 47 wherein the first functional group on the surface of the solid support is an epoxy group and the second functional group of the single-stranded nucleic acid probe is an amino group.

51. The probe array according to claim 42 wherein the spots are formed by supplying a liquid containing the probe on the solid support.

52. The probe array according to claim 42 wherein the probe is an oligopeptide or a polypeptide with a specific amino acid sequence.

53. The probe array according to claim 42 wherein the probe is a protein.

54. The probe array according to claim 53 wherein the protein is an antibody.

55. The probe array according to claim 53 wherein the protein is an enzyme.

56. The probe array according to claim 42 wherein the probe is an antigen.

57. The probe array according to claim 42 wherein a distance between the adjacent spots is not smaller than a maximum width of the spot.

58. The probe array according to claim 41 wherein the solid support is partitioned by a matrix arranged in a pattern on the surface, a plurality of wells whose bottom is the surface of the solid support exposed in a pattern are provided, and the liquid is supplied to the respective wells.

59. The probe array according to claim 58 wherein the probe is a single-stranded nucleic acid probe.

60. The probe array according to claim 59 wherein the single-stranded nucleic acid probe includes a single-stranded DNA probe.

61. The probe array according to claim 59 wherein the single-stranded nucleic acid includes a RNA probe.

62. The probe array according to claim 59 wherein the single-stranded nucleic acid includes a single-stranded PNA probe.

63. The probe array according to claim 62 wherein the single-stranded nucleic acid is covalently bound to the surface of the solid support by a reaction between the first functional group of the surface of the solid surface and the second functional group on the single-stranded nucleic acid probe.

64. The probe array according to claim 63 wherein the first functional group on the surface of the solid support is a maleimido group and the second functional group of the single-stranded nucleic acid probe is a thiol (SH) group.

65. The probe array according to claim 64 wherein the single-stranded nucleic acid probe is a single-stranded PNA probe and contains a cysteine residue on an N-terminal side.

66. The probe array according to claim 63 wherein the first functional group on the surface of the solid support is an epoxy group and the second functional group of the single-stranded nucleic acid probe is an amino group.

67. The probe array according to claim 58 wherein the spots are formed by supplying a liquid containing a probe on the solid support.

68. The probe array according to claim 58 wherein the probe is an oligopeptide or a polypeptide with a specific amino acid sequence.

69. The probe array according to claim 58 wherein the probe is a protein.

70. The probe array according to claim 69 wherein the protein is an antibody.

71. The probe array according to claim 69 wherein the protein is an enzyme.

72. The probe array according to claim 58 wherein the probe is an antigen.

73. The probe array according to claim 58 wherein the matrix is opaque.

74. The probe array according to claim 73 wherein the solid support is optically transparent.

75. The probe array according to claim 58 wherein the matrix comprises a resin.

76. The probe array according to claim 58 wherein the probe is attached only to the wells.

77. The probe array according to claim 58 wherein the matrix has a thickness of 1-20 μ

m.

78. The probe array according to claim 58 wherein the wells have a maximum width of 200 μ

m.

79. The probe array according to claim 58 wherein a distance between the wells is ½

-2 times the maximum width of the wells.

80. The probe array according to claim 41 wherein the probe array comprises at least 2 spots each of which comprises a different kind of probe.

81. A method of manufacturing a probe array having a plurality of spots arranged independently in a plurality of sites on a surface of a solid support, the spots containing a probe which can bind specifically to a target substance comprising a step of supplying a liquid containing the probe and attaching the liquid to a predetermined site on the surface of the solid support by means of an ink jet method to form the spots.

82. The method of manufacturing according to claim 81 wherein the probe is a single-stranded nucleic acid probe.

83. The method of manufacturing according to claim 82 wherein the single-stranded nucleic acid probe is a single-stranded DNA probe.

84. The method of manufacturing according to claim 82 wherein the single-stranded nucleic acid probe is an RNA probe.

85. The method of manufacturing according to claim 82 wherein the single-stranded nucleic acid probe is a single-stranded PNA probe.

86. The method of manufacturing according to claim 82 wherein the surface of the solid surface has a first functional group and the single-stranded nucleic acid probe has a second functional group, and the first and the second functional groups react each other by contact.

87. The method of manufacturing according to claim 86 wherein the first functional group on the surface of the solid support is a maleimido group and the second functional group of the single-stranded nucleic acid probe is a thiol (SH) group.

88. The method of manufacturing according to claim 87 wherein the solid support is a glass plate and a maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with N-(6-maleimidocaproyloxy) succinimide.

89. The method of manufacturing according to claim 87 wherein the solid support is a glass plate and the maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with succinimidyl-4-(maleimido phenyl) butyrate.

90. The method of manufacturing according to claim 87 wherein the maleimido group is reacted with the thiol group for at least 30 minutes.

91. The method of manufacturing according to claim 90 wherein the single-stranded nucleic acid is a single-stranded PNA probe, at the terminus of which the thiol group exists, the maleimido group is reacted with the thiol group for at least 2 hours.

92. The method of manufacturing according to claim 91 wherein the thiol group at the terminus of the single-stranded PNA probe is introduced by binding cysteine group to an N-terminal of the single-stranded PNA probe.

93. The method of manufacturing according to claim 86 wherein the first functional group on the surface of the solid support is an epoxy group and the second functional group of the single-stranded nucleic acid probe has is an amino group.

94. The method of manufacturing according to claim 93 wherein the solid support is a glass plate and the epoxy group is introduced by applying a silane compound having an epoxy group in the molecule thereof on the surface of the glass plate and reacting the compound with the glass plate.

95. The method of manufacturing according to claim 93 wherein the epoxy group is introduced by applying polyglycidyl methacrylate having an epoxy group on the solid support.

96. The method of manufacturing according to claim 93 wherein the liquid contains urea at 5-10 wt %, glycerin at 5-10 wt %, thiodiglycol at 5-10 wt %, and an acetylene alcohol at 1 wt % of the liquid.

97. The method of manufacturing according to claim 96 wherein the acetylene alcohol has a structure represented by the following general formula (I):

98. The method of manufacturing according to claim 82 wherein a concentration of the single-stranded nucleic acid probe in the liquid is 0.05-500 μ

M.

99. The method of manufacturing according to claim 98 wherein the concentration of the single-stranded nucleic acid probe in the liquid is 2-50 μ

M.

100. The method of manufacturing according to claim 82 wherein a length of the single-stranded nucleic acid probe is 2-5,000 bases.

101. The method of manufacturing according to claim 100 wherein the length of the single-stranded nucleic acid probe is 2-60 bases.

102. The method of manufacturing according to claim 81 wherein the ink jet method is a bubble jet method.

103. The method of manufacturing according to claim 81 wherein the liquid is supplied so as to form independent spots in a density of 10,000 spots per square inch on the solid support of higher.

104. The method of manufacturing according to claim 81 wherein the probe is an oligopeptide or a polypeptide with a specific amino acid sequence.

105. The method of manufacturing according to claim 81 wherein the probe is a protein.

106. The method of manufacturing according to claim 105 wherein the protein is an antibody.

107. The method of manufacturing according to claim 105 wherein the protein is an enzyme.

108. The method of manufacturing according to claim 81 wherein the probe is an antigen.

109. The method of manufacturing according to claim 81 wherein the solid support has a flat surface and homogenous surface properties.

110. The method of manufacturing according to claim 109 wherein blocking is performed, following spotting of the probe to the solid support, to prevent a sample from attaching to the surface other than the spots.

111. The method of manufacturing according to claim 110 wherein the blocking comprises a step of immersing the solid support to which the spots have been formed in an aqueous solution of bovine serum albumin.

112. The method of manufacturing according to claim 111 wherein a concentration of the aqueous solution of bovine serum albumin is 0.1-5%.

113. The method of manufacturing according to claim 111 wherein the solid support is immersed in an aqueous solution of bovine serum albumin for at least 2 hours.

114. The method of manufacturing according to claim 109 wherein the liquid is supplied on the surface of the solid support so as to obtain a distance between the adjacent spots not smaller than a maximum width of the spot.

115. The method of manufacturing according to claim 81 wherein the solid support is partitioned by a matrix arranged in a pattern on the surface, a plurality of wells whose bottom is the surface of the solid support exposed in a pattern are provided, and the liquid is supplied to the respective wells.

116. The method of manufacturing according to claim 115 wherein the solid support is optically transparent and the matrix is opaque.

117. The method of manufacturing according to claim 115 wherein the matrix comprises a resin.

118. The method of manufacturing according to claim 115 wherein the surface of the matrix is hydrophobic.

119. The method of manufacturing according to claim 115 wherein the bottom of the wells is hydrophilic.

120. The method of manufacturing according to claim 115 wherein the wells have a maximum width of 200 μ

m.

121. The method of manufacturing according to claim 115 wherein the matrix has a width ½

-2 times the maximum width of the wells.

122. The method of manufacturing according to claim 115 wherein a thickness of the matrix is 1-20 μ

m.

123. The method of manufacturing according to claim 115 wherein the matrix pattern is formed by photolithography.

124. The method of manufacturing according to claim 123 wherein the photolithography comprises a step of forming a resin layer on the surface of the solid support, forming a photoresist layer on the resin layer, exposing the photoresist layer to light in a pattern corresponding to the matrix pattern, and developing to form the pattern of the photoresist on the resin layer;

and a step of patterning the resin layer using the pattern of the photoresist as a mask and then removing the pattern of the photoresists.

125. The method of manufacturing according to claim 123 wherein the photolithography comprises the steps of forming a photosensitive resin layer on the surface of the solid support, exposing the photosensitive resin layer to light in a pattern corresponding to the matrix pattern, and developing.

126. The method of manufacturing according to claim 125 wherein the photosensitive resin layer is selected from the group consisted of a UV resist, a DEEP-UV resist, or an ultraviolet cure resin.

127. The method of manufacturing according to claim 126 wherein the UV resist is selected from the group consisted of a cyclized polyisoprene-aromatic bisazide resist, a phenol resin-aromatic azide compound resist, or a novolak resin-diazonaphtoquinone resist.

128. The method of manufacturing according to claim 126 wherein the DEEP-UV resin is a radiolysable polymer resist or a dissolution suppressant resist.

129. The method of manufacturing according to claim 128 wherein the radiation decomposition polymer resist is at least one selected from a group consisting of polymethyl methacrylate, polymethylene sulfone, polyhexafluorobutyl methacrylate, polymethylisopropenyl ketone, and poly-1-trimethylsilyl propyne bromide.

130. The method of manufacturing according to claim 128 wherein the dissolution suppressant resist is o-nitrobenzyl cholate ester.

131. The method of manufacturing according to claim 126 wherein the DEEP-UV resist is polyvinylphenol-3,3′

-diazidediphenyl sulfone or polyglycidyl polymethacrylate.

132. The method of manufacturing according to claim 125 wherein water repellency of the matrix pattern formed by patterning of the photosensitive resin layer is further improved by postbaking of the matrix pattern.

133. The method of manufacturing according to claim 115 wherein a first functional group which can form a covalent bond with a second functional group of the probe is introduced on the surface of the solid support prior to formation of the wells.

134. The method of manufacturing according to claim 115 wherein a first functional group which can form a covalent bond with a second functional group of the probe is introduced on the surface of the solid support following formation of the wells.

135. The method of manufacturing according to claim 134 wherein a solution containing a compound for introducing the first functional group to the surface of the solid support is supplied to the wells.

136. The method of manufacturing according to claim 135 wherein the solution is supplied to the wells by means of the ink jet method.

137. The method of manufacturing according to claim 136 wherein the solution is a silane coupling agent containing a silane compound having an epoxy group or an amino group in its molecule.

138. The method of manufacturing according to claim 136 wherein the solution contains a compound which can react with an amino group on a glass substrate to introduce a maleimido group on the glass substrate.

139. The method of manufacturing according to claim 138 wherein the compound is N-maleimidocaproyloxy succinimide or succinimidyl-4-(maleimidophenyl) butyrate.

140. A method for detecting whether a target substance is contained in a sample, comprising the steps of:

providing a probe array comprising a plurality of spots each containing a probe which specifically binds to the target substance, the spots being arranged independently on a solid support;

contacting the sample with each of the spots; and

detecting presence or absence of a reacted product between the target substance and the probe, wherein the respective spots are formed by spotting a liquid containing the probe on the solid support by an ink jet method.

141. The method according to claim 140 wherein the target substance is a single-stranded nucleic acid having a first base sequence and the probe is a single-stranded nucleic acid probe having a second base sequence complementary to the first base sequence.

142. The method according to claim 141 wherein the single-stranded nucleic acid probe is a single-stranded DNA probe.

143. The method according to claim 141 wherein the single-stranded nucleic acid probe is an RNA probe.

144. The method according to claim 141 wherein the single-stranded nucleic acid probe is a single-stranded PNA probe.

145. The method according to claim 141 wherein the surface of the solid surface has a first functional group and the single-stranded nucleic acid probe has a second functional group, respectively, and the functional groups react each other by contact.

146. The method according to claim 145 wherein the first functional group on the surface of the solid support is a maleimido group and the second functional group of the single-stranded nucleic acid probe is a thiol (SH) group.

147. The method according to claim 146 wherein the solid support is a glass plate and the maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with N-(6-maleimidocaproyloxy) succinimide.

148. The method according to claim 146 wherein the solid support is a glass plate and the maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with succinimidyl-4-(maleimidophenyl) butyrate.

149. The method according to claim 146 wherein the maleimido group is reacted with the thiol group for at least 30 minutes.

150. The method according to claim 149 wherein the single-stranded nucleic acid comprises a single-stranded PNA probe having a thiol group on the terminus and the maleimido group is reacted with the thiol group for at least 2 hours.

151. The method according to claim 146 wherein the thiol group at a terminus of the single-stranded PNA probe is introduced by binding cysteine group to an N-terminus of the single-stranded PNA probe.

152. The method according to claim 145 wherein the first functional group on the surface of the solid support is an epoxy group and the second functional group of the single-stranded nucleic acid probe is an amino group.

153. The method according to claim 152 wherein the solid support is a glass plate and the epoxy group is introduced by applying a silane compound having an epoxy group in the molecule thereof on the surface of the glass plate and reacting the compound with the glass plate.

154. The method according to claim 152 wherein the epoxy group is introduced by applying polyglycidyl methacrylate having an epoxy group on the solid support.

155. The method according to claim 141 wherein the liquid contains urea at 5-10 wt %, glycerin at 5-10 wt %, thiodiglycol at 5-10 wt %, and an acetylene alcohol at 1 wt % of the liquid.

156. The method according to claim 155 wherein the acetylene alcohol has a structure represented by the following general formula (I):

157. The method according to claim 155 wherein a concentration of the single-stranded nucleic acid probe in the liquid is 0.05-500 μ

M.

158. The method according to claim 157 wherein a concentration of the single-stranded nucleic acid probe in the liquid is 2-50 μ

M.

159. The method according to claim 155 wherein a length of the single-stranded nucleic acid probe is 2-5,000 bases.

160. The method according to claim 159 wherein a length of the single-stranded nucleic acid probe is 2-60 bases.

161. The method according to claim 141 wherein the ink jet method is a bubble jet method.

162. The method according to claim 140 wherein the probe is an oligopeptide or a polypeptide with a specific amino acid sequence.

163. The method according to claim 140 wherein the probe is a protein.

164. The method according to claim 163 wherein the protein is an antibody.

165. The method according to claim 163 wherein the protein is an enzyme.

166. The method according to claim 140 wherein the probe is an antigen.

167. The method according to claim 140 wherein the liquid is supplied so as to form independent spots in a density of 10,000 spots per square inch on the solid support.

168. The method according to claim 140 wherein the solid support has a flat surface and homogenous surface properties.

169. The method according to claim 168 wherein the liquid is supplied on the surface of the solid support so as to obtain a distance between the adjacent spots not smaller than the maximum width of the spots.

170. The method according to claim 168 wherein blocking is performed on the surface of the solid support to prevent the sample from attaching to the surface other than the spots of the surface of the solid support.

171. The method according to claim 170 wherein blocking is achieved by using bovine serum albumin.

172. The method according to claim 140 wherein the solid support is partitioned by a matrix arranged in a pattern on the surface, a plurality of wells whose bottom is the surface of the solid support exposed in the pattern are provided, and the liquid is supplied to the respective wells.

173. The method according to claim 172 wherein the solid support is optically transparent and the matrix is opaque.

174. The method according to claim 172 wherein the matrix comprises a resin.

175. The method according to claim 172 wherein the surface of the matrix is hydrophobic.

176. The method according to claim 172 wherein the bottom of the wells is hydrophilic.

177. The method according to claim 172 wherein the matrix has a thickness of 1-20 μ

m.

178. The method according to claim 172 wherein the wells have a maximum width of 200 μ

m.

179. The method according to claim 172 wherein the matrix has a width ½

-2 times the maximum width of the wells.

180. A method of identifying a structure of a target substance contained in a sample comprising the steps of:

preparing a probe array provided with spots of a probe, the probe being able to bind specifically to the target substance, on a surface of a solid support;

contacting the sample to the spots; and

detecting binding between the target substance and the probe.

181. The method of identification according to claim 180 wherein the target substance is a single-stranded nucleic acid, the structure to be identified is a base sequence of the single-stranded nucleic acid as the target substance,

the probe array is provided with a plurality of spots each of which contains single-stranded nucleic acids with different base sequences on a solid support, at least one of the spots contain a single-stranded nucleic acid with a base sequence complementary to that anticipated for the single-stranded nucleic acid as the target substance, and the plurality of spots are formed by attaching a liquid containing the respective single-stranded nucleic acids on the solid support by means of an ink jet method.

182. The method of identification according to claim 181 wherein the single-stranded nucleic acid probe is a single-stranded DNA probe.

183. The method of identification according to claim 181 wherein the single-stranded nucleic acid probe is an RNA probe.

184. The method of identification according to claim 181 wherein the single-stranded nucleic acid probe is a single-stranded PNA probe.

185. The method of identification according to claim 181 wherein the surface of the solid surface and the single-stranded nucleic acid probe have a first and a second functional groups, respectively, and the functional groups react each other by contact.

186. The method of identification according to claim 181 wherein the first functional group on the surface of the solid support is a maleimido group and the second functional group of the single-stranded nucleic acid probe is a thiol (SH) group.

187. The method of identification according to claim 186 wherein the solid support is a glass plate and the maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with N-(6-maleimidocaproyloxy) succinimide.

188. The method of identification according to claim 186 wherein the solid support is a glass plate and the maleimido group is introduced by introducing an amino group on the surface of the glass plate and then reacting the amino group with succinimidyl-4-(maleimido phenyl) butyrate.

189. The method of identification according to claim 186 wherein the maleimido group is reacted with the thiol group for at least 30 minutes.

190. The method of identification according to claim 189 wherein the single-stranded nucleic acid is a single-stranded PNA probe having a thiol group on the terminus thereof and the maleimido group is reacted with the thiol group for at least 2 hours.

191. The method of identification according to claim 186 wherein the thiol group at the terminus of the single-stranded PNA probe is introduced by binding cysteine group to an N-terminus of the single-stranded PNA probe.

192. The method of identification according to claim 185 wherein the first functional group on the surface of the solid support is an epoxy group and the second functional group of the single-stranded nucleic acid probe is an amino group.

193. The method of identification according to claim 192 wherein the solid support is a glass plate and the epoxy group is introduced by applying a silane compound having an epoxy group in the molecule on the surface of the glass plate and reacting the compound with the glass plate.

194. The method of identification according to claim 192 wherein the epoxy group is introduced by applying polyglycidyl methacrylate having an epoxy group on the solid support.

195. The method of identification according to claim 181 wherein the liquid contains urea at 5-10 wt %, glycerin at 5-10 wt %, thiodiglycol at 5-10 wt %, and acetylene alcohol at 1 wt % of the liquid.

196. The method of identification according to claim 195 wherein the acetylene alcohol has a structure represented by the following general formula (I):

197. The method of identification according to claim 195 wherein a concentration of the single-stranded nucleic acid probe in the liquid is 0.05-500 μ

M.

198. The method of identification according to claim 197 wherein the concentration of the single-stranded nucleic acid probe in the liquid is 2-50 μ

M.

199. The method of identification according to claim 197 wherein a length of the single-stranded nucleic acid probe is 2-5,000 bases.

200. The method of identification according to claim 199 wherein a length of the single-stranded nucleic acid probe is 2-60 bases.

201. The method of identification according to claim 181 wherein the ink jet method is a bubble jet method.

202. The method of identification according to claim 180 wherein the probe is an oligopeptide or a polypeptide with a specific amino acid sequence.

203. The method of identification according to claim 180 wherein the probe is a protein.

204. The method of identification according to claim 203 wherein the protein is an antibody.

205. The method of identification according to claim 203 wherein the protein is an enzyme.

206. The method of identification according to claim 180 wherein the probe is an antigen.

207. The method of identification according to claim 180 wherein the liquid is supplied so as to form independent spots in a density of 10,000 spots per square inch on the solid support.

208. The method of identification according to claim 180 wherein the solid support has a flat surface and homogenous surface properties.

209. The method of identification according to claim 208 wherein the liquid is supplied on the surface of the solid support so as to obtain a distance between the adjacent spots not smaller than the maximum width of the spots.

210. The method of identification according to claim 208 wherein blocking is performed on the surface of the solid support to prevent the sample from attaching to the surface other than spots of the surface of the solid support.

211. The method of identification according to claim 210 wherein blocking is achieved by using bovine serum albumin.

212. The method of identification according to claim 180 wherein the solid support is partitioned by a matrix arranged in a pattern on the surface, a plurality of wells whose bottom is the surface of the solid support exposed in the pattern are provided, and the liquid is supplied to the respective wells.

213. The method of identification according to claim 212 wherein the solid support is optically transparent and the matrix is opaque.

214. The method of identification according to claim 212 wherein the matrix comprises a resin.

215. The method of identification according to claim 212 wherein the surface of the matrix is hydrophobic.

216. The method of identification according to claim 212 wherein the bottom of the wells is hydrophilic.

217. The method of identification according to claim 212 wherein the matrix has a thickness of 1-20 μ

m.

218. The method of identification according to claim 212 wherein the wells have a maximum width of 200 μ

m.

219. The method of identification according to claim 212 wherein the matrix has a width ½

-2 times a maximum width of the wells.