System, array and non-porous solid support comprising fixed or immobilized nucleic acids

DC CAFC

US 7,064,197 B1
Filed: 06/07/1995
Issued: 06/20/2006
Est. Priority Date: 01/27/1983
Status: Expired due to Term

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1. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein at least one single-stranded nucleic acid is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).

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Abstract

Nucleic acids are fixed or immobilized to non-porous solid supports (substrates), and include systems containing such supports and arrays with fixed or immobilized nucleic acids. These compositions are useful for nucleic acid analyses and a host of applications, including, for example, detection, mutational analysis and quantification. The non-porous solid supports can be transparent or translucent, and the surfaces can be treated with agents to fix or immobilize the nucleic acids. Such agents include, for example, amine providing compounds, epoxy compounds and acid solutions. The fixed or immobilized nucleic acids can be unlabeled, or labeled with at least one non-radioactive signaling moiety, such as the case when the nucleic acids are double-stranded.

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238 Claims

1. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein at least one single-stranded nucleic acid is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).
- View Dependent Claims (16, 23, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 61, 62, 63, 64, 102, 103, 104, 236)
- - 16. The non-porous solid support of claims 1, 2, 12, 13, 14, 15 or 4, wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.
  - 23. The non-porous solid support of claims 1, 2, 12, 13, 14, 15, 3, 4 or 5, wherein said non-porous solid support comprises an arrangement of wells, tubes or cuvettes.
  - 31. The non-porous solid support of claim 1, wherein said nucleic acid comprises a nucleic acid sequence complementary to a nucleic acid sequence of interest sought to be identified, quantified or sequenced.
  - 32. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said non-porous solid support comprises glass or plastic.
  - 33. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said non-porous solid support comprises a plate or plates, a well or wells, a microtiter well or microtiter wells, a depression or depressions, a tube or tubes, or a cuvette or cuvettes.
  - 34. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said non-porous solid support has been treated with a surface treatment agent, a blocking agent, or both.
  - 35. The non-porous solid support of claim 34, wherein said surface treatment agent comprises an amine providing compound, an epoxy glue or solution, an acid solution, or ammonium acetate.
  - 36. The non-porous solid support of claim 34, wherein said blocking agent comprises Denhardt'"'"'s solution.
  - 37. The non-porous solid support of claim 35, wherein said acid compound comprises nitric acid.
  - 38. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said fixation or immobilization to said non-porous solid support is covalent.
  - 39. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said non-porous solid support is transparent or translucent.
  - 40. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said non-porous solid support comprises a plate or plates.
  - 41. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said non-porous solid support comprises a well or wells, a microtiter well or microtiter wells, or a depression or depressions.
  - 42. The non-porous solid support of claim 35, wherein said amine providing compound comprises dodecadiamine (DDA), polylysine (PPL) or 6-aminohexane.
  - 61. The non-porous solid support of claims 1 or 4, wherein said nucleic acid is DNA.
  - 62. The non-porous solid support of claims 1 or 4, wherein said single-stranded nucleic acid is unlabeled.
  - 63. The non-porous solid support of claims 1 or 4, comprising more than one single-stranded nucleic acid.
  - 64. The non-porous solid support of claims 1 or 4, wherein said nucleic acid is RNA.
  - 102. The system of claims 7 or 16, wherein one strand of said at least one double-stranded nucleic acid comprises a nucleic acid sequence of interest sought to be identified, quantified or sequenced.
  - 103. The system of claims 7 or 16, wherein said non-radioactive signaling moiety is quantifiable in or from a fluid or solution, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.
  - 104. The system of claims 7 or 16, wherein said non-porous solid support is transparent or translucent, and said non-radioactive signaling moiety is quantifiable in or from a fluid or solution or from said non-porous solid support, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.
  - 236. The non-porous solid support of claims 1, 2, 3, 4 or 5, wherein said fixation or immobilization to said non-porous solid support is non-covalent.

2. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein at least one double-stranded nucleic acid is fixed or immobilized to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s), and wherein one nucleic acid strand of said at least one double-stranded nucleic acid has covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety.
- View Dependent Claims (43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 65, 66, 67, 228, 229)
- - 43. The non-porous solid support of claims 2, 3 or 5, wherein one strand of said at least one double-stranded nucleic acid is fixed or immobilized to said non-porous solid support and the other strand of said at least one double-stranded nucleic acid is hybridized to said one strand.
  - 44. The non-porous solid support of claims 2, 3 or 5, wherein said nucleic acid is DNA.
  - 45. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety comprises a chromagen or a chromagenic compound.
  - 46. The non-porous solid support of claims 2, 3 or 5, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by photometric techniques.
  - 47. The non-porous solid support of claims 2, 3 or 5, comprising more than one double-stranded nucleic acid.
  - 48. The non-porous solid support of claims 2, 3 or 5, wherein said nucleic acid is RNA.
  - 49. The non-porous solid support of claims 2, 3 or 5, wherein said nucleic acid comprises both DNA and RNA.
  - 50. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety is a colored dye compound.
  - 51. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety is a fluorogen or a fluorescent compound.
  - 52. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety is a chemiluminescent compound.
  - 53. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety comprises a chelating compound.
  - 54. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety comprises an enzyme or an enzymatic compound.
  - 55. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety comprises a coenzyme.
  - 56. The non-porous solid support of claims 2, 3 or 5, wherein said non-radioactive signaling moiety comprises a biotin.
  - 57. The non-porous solid support of claims 2, 3 or 5, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by spectrophotometric techniques.
  - 58. The non-porous solid support of claims 2, 3 or 5, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by colorimetric techniques.
  - 59. The non-porous solid support of claims 2, 3 or 5, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by fluorometric techniques.
  - 60. The non-porous solid support of claims 2, 3 or 5, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by chemiluminescent techniques.
  - 65. The non-porous solid support of claims 2 or 3, wherein one strand of said at least one double-stranded nucleic acid comprises a nucleic acid sequence of interest sought to be identified, quantified or sequenced.
  - 66. The non-porous solid support of claims 2 or 3, wherein said non-radioactive signaling moiety is quantifiable in or from a fluid or solution, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.
  - 67. The non-porous solid support of claims 2 or 3, wherein said non-porous solid support is transparent or translucent, and said non-radioactive signaling moiety is quantifiable in or from a fluid or solution or from said non-porous solid support, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.
  - 228. The non-porous solid support of claims 2, 29, 30, 3 or 5, wherein said bridging moiety comprises biotin-avidin, biotin-streptavidin or sugar-lectin.
  - 229. The non-porous solid support of claims 2, 29, 30, 3 or 5, wherein one strand of said at least one double-stranded nucleic acid is fixed or immobilized to said non-porous solid support by sandwich hybridization.

3. A non-porous solid support comprising at least one double-stranded nucleic acid fixed or immobilized thereto, wherein one nucleic acid strand of said at least one double-stranded nucleic acid has covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety,and wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.
- View Dependent Claims (24, 235)
- - 24. The non-porous solid support of claim 3 or 5, comprising one or more amines.
  - 235. The non-porous solid support of claim 3 or 5, wherein said non-porous solid support comprises one or more epoxides.

4. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, and at least one nucleic acid strand or sequence fixed or immobilized to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s), and wherein another nucleic acid strand or sequence is hybridized to said at least one nucleic acid strand or sequence, said another nucleic acid strand or sequence having covalently attached thereto either:
- (l) a non-radioactive signaling moiety, or(ll) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety.

5. A non-porous solid support comprising at least one nucleic acid strand or sequence fixed or immobilized to said non-porous solid support, and wherein one other nucleic acid strand or sequence is hybridized to said at least one nucleic acid strand or sequence, said other nucleic acid strand or sequence having covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety,and wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.

6. A system comprising a non-porous solid support which comprises one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein at least one single-stranded nucleic acid is fixed or immobilized thereto in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).
- View Dependent Claims (68, 69, 70, 100, 101, 230)
- - 68. The system of claim 6, wherein said nucleic acid comprises a nucleic acid sequence complementary to a nucleic acid sequence of interest sought to be identified, quantified or sequenced.
  - 69. The system of claim 6, wherein said single-stranded nucleic acid is unlabeled.
  - 70. The system of claim 6, comprising more than one single-stranded nucleic acid.
  - 100. The system of claims 6 or 9, wherein said nucleic acid is DNA.
  - 101. The system of claims 6 or 9, wherein said nucleic acid is RNA.
  - 230. The system of claims 6, 7, 8 or 9, wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.

7. A system comprising a non-porous solid support which comprises one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein at least one double-stranded nucleic acid is fixed or immobilized to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s), and wherein one nucleic acid strand of said double-stranded nucleic acid has covalently attached thereto either:
- (l) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety.
- View Dependent Claims (71)
- - 71. The system of claim 7, wherein said bridging moiety comprises biotin-avidin, biotin-streptavidin or sugar-lectin.

8. A system comprising a non-porous solid support which comprises one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein DNA or RNA is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).

9. A system comprising a non-porous solid support which comprises one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein nucleic acid is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).

10. A system comprising a non-porous solid support which comprises double-stranded nucleic acid fixed or immobilized thereto, wherein one nucleic acid strand of said double-stranded nucleic acid has covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety,and wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.
- View Dependent Claims (72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 227, 231, 232, 233, 238)
- - 72. The system of claims 6, 7, 8, 9, 10 or 11, wherein said non-porous solid support comprises glass or plastic.
  - 73. The system of claims 6, 7, 8, 9, 10, or 11, wherein said non-porous solid support comprises a plate or plates, a well or wells, a microtiter well or microtiter wells, a depression or depressions, a tube or tubes, or a cuvette or cuvettes.
  - 74. The system of claims 6, 7, 8, 9, 10 or 11, wherein said non-porous solid support has been treated with a surface treatment agent, a blocking agent, or both.
  - 75. The system of claim 74, wherein said surface treatment agent comprises an amine providing compound, an epoxy glue or solution, an acid solution or ammonium acetate.
  - 76. The system of claim 75, wherein said acid compound comprises nitric acid.
  - 77. The system of claim 74, wherein said blocking agent comprises Denhardt'"'"'s solution.
  - 78. The system of claims 6, 7, 8, 8, 10 or 11, wherein said fixation or immobilization to said non-porous solid support is covalent.
  - 79. The system of claims 6, 7, 8, 9, 10 or 11, wherein said fixation or immobilization to said non-porous solid support is non-covalent.
  - 80. The system of claims 6, 7, 8, 9, 10 or 11 wherein said non-porous solid support is transparent or translucent.
  - 81. The system of claims 7, 10 or 11, wherein one strand of said double-stranded nucleic acid is fixed or immobilized to said non-porous solid support and the other nucleic acid strand of said double-stranded nucleic acid is hybridized to said one strand.
  - 82. The system of claims 7, 10 or 11, wherein said nucleic acid is DNA.
  - 83. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises a chromagen or a chromagenic compound.
  - 84. The system of claims 7, 10 or 11, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by photometric techniques.
  - 85. The system of claims 7, 10 or 11, comprising more than one double-stranded nucleic acid.
  - 86. The system of claims 7, 10 or 11, wherein one strand of said double-stranded nucleic acid is fixed or immobilized to said non-porous solid support and the other strand of said double-stranded nucleic acid is hybridized to said one strand.
  - 87. The system of claims 7, 10 or 11, wherein said nucleic acid comprises RNA.
  - 88. The system of claims 7, 10 or 11, wherein said nucleic acid comprises both DNA and RNA.
  - 89. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises a colored dye compound.
  - 90. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises a fluorogen or a fluorescent compound.
  - 91. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises a chemiluminescent compound.
  - 92. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises a chelating compound.
  - 93. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises an enzyme or an enzymatic compound.
  - 94. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises a coenzyme.
  - 95. The system of claims 7, 10 or 11, wherein said non-radioactive signaling moiety comprises a biotin.
  - 96. The system of claims 7, 10 or 11, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by spectrophotometric techniques.
  - 97. The system of claims 7, 10 or 11, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by colorimetric techniques.
  - 98. The system of claims 7, 10 or 11, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by fluorometric techniques.
  - 99. The system of claims 7, 10 or 11, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by chemiluminescent techniques.
  - 227. The system of claims 8, 9, 10 or 11, wherein said non-porous solid support comprises glass or plastic.
  - 231. The system of claims 6, 7, 8, 9, 10 or 11, wherein said non-porous solid support comprises an arrangement of wells, tubes or cuvettes.
  - 232. The system of claims 6, 7, 8, 9, 10 or 11, wherein said non-porous solid support comprises a plate or plates.
  - 233. The system of claims 6, 7, 8, 9, 10 or 11, wherein said non-porous solid support comprises a well or wells, a microtiter well or microtiter wells, or a depression or depressions.
  - 238. The system of claim 75, wherein said amine providing compound comprises dodecadiamine (DDA), polylysine (PPL) or 6-aminohexane.

11. A system comprising a non-porous solid support which comprises a double-stranded nucleic acid fixed or immobilized thereto, wherein one nucleic acid strand of said double-stranded nucleic acid has covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety,and wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.

12. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein single-stranded nucleic acid is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).
- View Dependent Claims (226)
- - 226. The non-porous solid support of claims 12, 13, 14, 15, 29 or 30, wherein said non-porous solid support comprises glass or plastic.

13. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein a single-stranded nucleic acid is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).

14. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein nucleic acid is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).

15. A non-porous solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein DNA or RNA is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s).

17. An array comprising various single-stranded nucleic acids fixed or immobilized in hybridizable form to a non-porous solid support.
- View Dependent Claims (105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 128, 129, 130, 131)
- - 105. The array of claims 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support comprises glass or plastic.
  - 106. The array of claims 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support comprises a plate or plates, a well or wells, a microtiter well or microtiter wells, a depression or depressions, a tube or tubes, or a cuvette or cuvettes.
  - 107. The array of claims 17, 18, 19, 20, 21 or 22, comprising one or more amines.
  - 108. The array of claims 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support has been treated with a surface treatment agent, a blocking agent, or both.
  - 109. The array of claim 108, wherein said surface treatment agent comprises an amine providing compound, an epoxy glue or solution, an acid solution or ammonium acetate.
  - 110. The array of claim 109, wherein said amine providing compound comprises dodecadiamine (DDA), polylysine (PPL) or 6-aminohexane.
  - 111. The array of claim 109, wherein said acid compound comprises nitric acid.
  - 112. The array of claim 108, wherein said blocking agent comprises Denhardt'"'"'s solution.
  - 113. The array of claims 17, 18, 19, 20, 21 or 22, wherein said fixation or immobilization to said non-porous solid support is covalent.
  - 114. The array of claims 17, 18, 19, 20, 21 or 22, wherein said fixation or immobilization to said non-porous solid support is non-covalent.
  - 115. The array of claims 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support is transparent or translucent.
  - 116. The array of claims 17, 18, 19, 20, 21 or 22, wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.
  - 117. The array of claims 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support comprises an arrangement of wells, tubes or cuvettes.
  - 118. The array of claims 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support comprises a plate or plates.
  - 119. The array of claims 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support comprises a well or wells, a microtiter well or microtiter wells, or a depression or depressions.
  - 120. The array of claim 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support comprises one or more hydroxyls.
  - 121. The array of claim 17, 18, 19, 20, 21 or 22, wherein said non-porous solid support comprises one or more epoxides.
  - 128. The array of claims 17 or 19, wherein said nucleic acids is DNA.
  - 129. The array of claims 17 or 19, wherein said single-stranded nucleic acids are unlabeled.
  - 130. The array of claims 17 or 19, wherein said nucleic acids is RNA.
  - 131. The array of claims 17 or 23, wherein said nucleic acids comprise nucleic acid sequences complementary to nucleic acid sequences of interest sought to be identified, quantified or sequenced.

18. An array comprising various double-stranded nucleic acids fixed or immobilized to a non-porous solid support, wherein at least one nucleic acid strand of said various double-stranded nucleic acids comprises at least one non-radioactive signaling moiety which is quantifiable or detectable.
- View Dependent Claims (122, 123, 124, 125, 126, 127, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149)
- - 122. The array of claims 18 or 20, wherein the first strands of said double-stranded nucleic acids are fixed or immobilized to said non-porous solid support and the other nucleic acid strands of said double-stranded nucleic acids are hybridized to said first strands.
  - 123. The array of claims 18 or 20, wherein said nucleic acids is DNA.
  - 124. The array of claims 18 or 20, wherein the first strands of said double-stranded nucleic acids are fixed or immobilized to said non-porous solid support and the other nucleic acid strands of said double-stranded nucleic acids are hybridized to said first strands.
  - 125. The array of claims 18 or 20, wherein one strand of said at least one double-stranded nucleic acid is fixed or immobilized to said non-porous solid support by sandwich hybridization.
  - 126. The array of claims 18 or 20, wherein said nucleic acids is RNA.
  - 127. The array of claims 18 or 02, wherein said nucleic acids comprises both DNA and RNA.
  - 132. The array of claims 18 or 24, wherein one strand of said double-stranded nucleic acids comprises a nucleic acid sequence of interest sought to be identified, quantified or sequenced.
  - 133. The array of claims 18 or 24, wherein said non-radioactive signaling moiety is quantifiable in or from a fluid or solution, said quantity being proportional to the amount or quantity of said label or labels.
  - 134. The array of claims 18 or 42, wherein said non-porous solid support is transparent or translucent, and said non-radioactive signaling moiety is quantifiable in or from a fluid or solution or from said non-porous solid support, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.
  - 135. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety comprises a chromagen or a chromagenic compound.
  - 136. The array of claims 18, 20, 21 or 22, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by photometric techniques.
  - 137. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is attached to said at least one nucleic acid strand through a bridging moiety.
  - 138. The array of claim 137, wherein said bridging moiety comprises biotin-avidin, biotin-streptavidin or sugar-lectin.
  - 139. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is a colored dye compound.
  - 140. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is a fluorogen or a fluorescent compound.
  - 141. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is a chemiluminescent compound.
  - 142. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is a chelating compound.
  - 143. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is an enzyme or an enzymatic compound.
  - 144. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is a coenzyme.
  - 145. The array of claims 18, 20, 21 or 22, wherein said non-radioactive signaling moiety is biotin.
  - 146. The array of claims 18, 20, 21 or 22, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by spectrophotometric techniques.
  - 147. The array of claims 18, 20, 21 or 22, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by colorimetric techniques.
  - 148. The array of claims 18, 20, 21 or 22, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by fluorometric techniques.
  - 149. The array of claims 18, 20, 21 or 22, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by chemiluminescent techniques.

19. An array comprising single-stranded nucleic acids fixed or immobilized in hybridizable form to a non-porous solid support.

20. An array comprising double-stranded nucleic acids fixed or immobilized to a non-porous solid support, wherein at least one nucleic acid strand of said double-stranded nucleic acids comprises at least one non-radioactive signaling moiety which is quantifiable or detectable.

21. An array comprising various nucleic acids, wherein single-stranded nucleic acids or sequences are fixed or immobilized to a non-porous solid support, and other single-stranded nucleic acids or sequences are hybridized to said fixed or immobilized single-stranded nucleic acids or sequences, said other nucleic acid strands or sequences comprising at least one non-radioactive signaling moiety which is quantifiable or detectable.

22. An array comprising various first single-stranded nucleic acids fixed or immobilized to a non-porous solid support, and further comprising second nucleic acids hybridized to said fixed or immobilized first nucleic acids, wherein said second nucleic acids comprise at least one non-radioactive signalling moiety which is quantifiable or detectable.
- View Dependent Claims (234)
- - 234. The non-porous solid support of claim 22 or 5, wherein said non-porous solid support comprises one or more hydroxyls.

25. An array comprising various single-stranded nucleic acids fixed or immobilized in hybridizable form to a non-porous solid support having wells or depressions.
- View Dependent Claims (150, 151, 152, 153, 154, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190)
- - 150. The array of claim 25, wherein said nucleic acids are DNA.
  - 151. The array of claim 25, wherein said nucleic acids comprise a gene sequence or pathogen sequence.
  - 152. The array of claim 25, wherein said single-stranded nucleic acids are unlabeled.
  - 153. The array of claims 25 or 26, wherein said non-porous solid support is transparent or translucent.
  - 154. The array of claim 25, wherein said nucleic acids are RNA.
  - 178. The array of claims 25 or 26, wherein said non-porous solid support comprises glass or plastic.
  - 179. The array of claims 25 or 26, wherein said non-porous solid support comprises one or more amines.
  - 180. The array of claims 25 or 26, wherein said non-porous solid support has been treated with a surface treatment agent, a blocking agent, or both.
  - 181. The array of claim 180, wherein said surface treatment agent comprises an amine providing compound, an epoxy glue or solution, an acid solution or ammonium acetate.
  - 182. The array of claim 181, wherein said amine providing compound comprises dodecadiamine (DDA), polylysine (PPL) or 6-aminohexane.
  - 183. The array of claim 181, wherein said acid compound comprises nitric acid.
  - 184. The array of claim 180, wherein said blocking agent comprises Denhardt'"'"'s solution.
  - 185. The array of claims 25 or 26, wherein said fixation or immobilization to said non-porous solid support is covalent.
  - 186. The array of claims 25 or 26, wherein said fixation or immobilization to said non-porous solid support is non-covalent.
  - 187. The array of claims 25 or 26, wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.
  - 188. The array of claims 25 or 26, wherein said non-porous solid support comprises an arrangement of said wells.
  - 189. The array of claim 25 or 26, wherein said non-porous solid support comprises one or more hydroxyls.
  - 190. The array of claim 25 or 26, wherein said non-porous solid support comprises one or more epoxides.

26. An array comprising various double-stranded nucleic acids fixed or immobilized to a non-porous solid support having wells or depressions, wherein at least one nucleic acid strand of each of said various double-stranded nucleic acids comprises at least one non-radioactive signaling moiety which is quantifiable or detectable.
- View Dependent Claims (155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177)
- - 155. The array of claim 26, wherein the first strands of said double-stranded nucleic acids are fixed or immobilized to said non-porous solid support and the other nucleic acid strands of said double-stranded nucleic acids are hybridized to said first strands.
  - 156. The array of claim 26, wherein said nucleic acids are DNA.
  - 157. The array of claim 26, wherein one strand of said double-stranded nucleic acids comprises a gene sequence or pathogen sequence.
  - 158. The array of claim 26, wherein said non-radioactive signaling moiety is quantifiable in or from a fluid or solution, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.
  - 159. The array of claim 26, wherein said non-porous solid support is transparent or translucent, and said non-radioactive signaling moiety is quantifiable in or from a fluid or solution or from said non-porous solid support, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.
  - 160. The array of claim 26, wherein said non-radioactive signaling moiety comprises a chromagen or a chromagenic compound.
  - 161. The array of claim 26, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by photometric techniques.
  - 162. The array of claim 26, wherein said at least one non-radioactive signaling moiety is attached to said at least one nucleic acid strand through a bridging moiety.
  - 163. The array of claim 162, wherein said bridging moiety comprises biotin-avidin, biotin-streptavidin or sugar-lectin.
  - 164. The array of claim 26, wherein one strand of various double-stranded nucleic acids is fixed or immobilized to said non-porous solid support by sandwich hybridization.
  - 165. The array of claim 26, wherein said nucleic acids are RNA.
  - 166. The array of claim 26, wherein said nucleic acids comprise both DNA and RNA.
  - 167. The array of claim 26, wherein said non-radioactive signaling moiety comprises a colored dye compound.
  - 168. The array of claim 26, wherein said non-radioactive signaling moiety comprises a fluorescent compound.
  - 169. The array of claim 26, wherein said non-radioactive signaling moiety comprises a chemiluminescent compound.
  - 170. The array of claim 26, wherein said non-radioactive signaling moiety comprises a chelating compound.
  - 171. The array of claim 26, wherein said non-radioactive signaling moiety comprises an enzyme or an enzymatic compound.
  - 172. The array of claim 26, wherein said non-radioactive signaling moiety comprises a coenzyme.
  - 173. The array of claim 26, wherein said non-radioactive signaling moiety comprises a biotin.
  - 174. The array of claim 26, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by spectrophotometric techniques.
  - 175. The array of claim 26, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by colorimetric techniques.
  - 176. The array of claim 26, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by fluorometric techniques.
  - 177. The array of claim 26, wherein a non-radioactive signal from said non-radioactive signaling moiety is quantifiable or detectable by chemiluminescent techniques.

27. A non-porous glass or non-porous plastic solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein at least one single-stranded nucleic acid is fixed or immobilized in hybridizable form to said non-porous solid support via said one or more amine(s), hydroxyl(s) or epoxide(s).
- View Dependent Claims (191, 192, 193, 194, 195, 212, 213, 214, 215, 216, 217, 218, 219, 221, 222, 223, 224, 225)
- - 191. The non-porous glass or non-porous plastic solid support of claim 27, wherein said nucleic acid is DNA.
  - 192. The non-porous glass or non-porous plastic solid support of claim 27, wherein said nucleic acid comprises a nucleic acid sequence complementary to a nucleic acid sequence of interest sought to be identified, quantified or sequenced.
  - 193. The non-porous glass or non-porous plastic solid support of claim 27, wherein said single-stranded nucleic acid is unlabeled.
  - 194. The non-porous glass or non-porous plastic solid support of claim 27, comprising more than one single-stranded nucleic acid fixed or immobilized in hybridizable form to said non-porous glass or non-porous plastic solid support.
  - 195. The non-porous glass or non-porous plastic solid support of claim 27, wherein said nucleic acid is RNA.
  - 212. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said non-porous glass or non-porous plastic solid support comprises a plate or plates, a well or wells, a microtiter well or microtiter wells, a depression or depressions, a tube or tubes, or a cuvette or cuvettes.
  - 213. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said non-porous glass or non-porous plastic solid support has been treated with a surface treatment agent, a blocking agent, or both.
  - 214. The non-porous glass or non-porous plastic solid support of claim 213, wherein said surface treatment agent comprises an amine providing compound, an epoxy glue or solution, an acid solution or ammonium acetate.
  - 215. The non-porous glass or non-porous plastic solid support of claim 214, wherein said amine providing compound comprises dodecadiamine (DDA), polylysine (PPL) or 6-aminohexane.
  - 216. The non-porous glass or non-porous plastic solid support of claim 214, wherein said acid compound comprises nitric acid.
  - 217. The non-porous glass or non-porous plastic solid support of claim 213, wherein said blocking agent comprises Denhardt'"'"'s solution.
  - 218. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said fixation or immobilization to said non-porous glass or non-porous plastic solid support is covalent.
  - 219. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said fixation or immobilization to said non-porous glass or non-porous plastic solid support is non-covalent.
  - 221. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said non-porous glass or non-porous plastic solid support is transparent or translucent.
  - 222. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.
  - 223. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said non-porous solid support comprises an arrangement of wells, tubes or cuvettes.
  - 224. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said non-porous glass or non-porous plastic solid support comprises a plate or plates.
  - 225. The non-porous glass or non-porous plastic solid support of claims 27 or 28, wherein said non-porous glass or non-porous plastic solid support comprises a well or wells, a microtiter well or microtiter wells, or a depression or depressions.

28. A non-porous glass or non-porous plastic solid support comprising one or more amine(s), hydroxyl(s) or epoxide(s) thereon, wherein at least one double-stranded nucleic acid is fixed or immobilized to said non-porous glass or non-porous plastic solid support via said one or more amine(s), hydroxyl(s) or epoxide(s), and wherein one nucleic acid strand of said double-stranded nucleic acid has covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety.
- View Dependent Claims (196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 220, 237)
- - 196. The non-porous glass or non-porous plastic solid support of claim 28, wherein said nucleic acid is DNA.
  - 197. The non-porous glass or non-porous plastic solid support of claim 28, wherein one strand of said at least one double-stranded nucleic acid comprises a nucleic acid sequence of interest sought to be identified, quantified or sequenced.
  - 198. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety is quantifiable in or from a fluid or solution, said quantity being proportional to the amount or quantity of said label or labels.
  - 199. The non-porous glass or non-porous plastic solid support of claim 28, comprising more than one double-stranded nucleic acid fixed or immobilized in hybridizable form to said non-porous glass or non-porous plastic solid support.
  - 200. The non-porous glass or non-porous plastic solid support of claim 28, wherein said bridging moiety comprises biotin-avidin, biotin-streptavidin or sugar-lectin.
  - 201. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises a chromagen or a chromagenic compound.
  - 202. The non-porous glass or non-porous plastic solid support of claim 28, wherein one strand of said at least one double-stranded nucleic acid is fixed or immobilized to said non-porous solid support by sandwich hybridization.
  - 203. The non-porous glass or non-porous plastic solid support of claim 28, wherein said nucleic acid is RNA.
  - 204. The non-porous glass or non-porous plastic solid support of claim 28, wherein said nucleic acid comprises both DNA and RNA.
  - 205. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises a colored dye compound.
  - 206. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises a fluorescent compound.
  - 207. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises a chemiluminescent compound.
  - 208. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises a chelating compound.
  - 209. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises an enzyme or an enzymatic compound.
  - 210. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises a coenzyme.
  - 211. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-radioactive signaling moiety comprises a biotin.
  - 220. The non-porous glass or non-porous plastic solid support of claim 28, wherein one strand of said double-stranded nucleic acid is fixed or immobilized to said non-porous glass or non-porous plastic solid support by sandwich hybridization or by hybridization to a complementary nucleic acid strand or sequence.
  - 237. The non-porous glass or non-porous plastic solid support of claim 28, wherein said non-porous glass or non-porous plastic solid support is transparent or translucent, and non-radioactive signaling moiety is quantifiable in or from a fluid or solution or from said non-porous glass or non-porous plastic solid support, said quantity being proportional to the amount or quantity of said non-radioactive signaling moiety.

29. A non-porous solid support comprising double-stranded nucleic acid fixed or immobilized thereto, wherein one nucleic acid strand of said double-stranded nucleic acid has covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety,wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.

30. A non-porous solid support comprising a double-stranded nucleic acid fixed or immobilized thereto, wherein one nucleic acid strand of said double-stranded nucleic acid has covalently attached thereto either:
- (i) a non-radioactive signaling moiety, or(ii) a non-radioactive bridging moiety covalently or non-covalently attached to a non-radioactive signaling moiety,wherein said fixation or immobilization is not to a cell fixed in situ to said non-porous solid support.

Specification

Resources

Litigation Campaign Assessment

Litigation Data

Current Assignee
Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
Original Assignee
Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
Inventors
Kirtikar, Dollie, Johnston, Kenneth H., Thalenfeld, Barbara E., Stavrianopoulos, Jannis G., Rabbani, Elazar
Primary Examiner(s)
Brusca, John S.

Application Number

US08/486,070
Time in Patent Office

4,031 Days
Field of Search

435/6, 435/287, 435/810, 435/283.1, 435/285.1, 435/287.1, 435/287.2, 435/287.7, 435/287.9, 435/288.7, 435/289.1, 435/297.1, 435/299.1, 436/501, 536/22.1, 935/78, 935/77, 935/88, 422/50, 422/55, 422/56, 422/57, 422/68.1, 422/69, 422/82.05, 422/82.06, 422/82.07, 422/82.08
US Class Current

536/24.3
CPC Class Codes

C07H 21/00   Compounds containing two or...

C12Q 1/6816   characterised by the detect...

C12Q 1/6834   Enzymatic or biochemical co...

G01N 33/535   with enzyme label or co-enz...

G01N 33/552   Glass or silica

G01N 33/581   with enzyme label (includin...

Y10S 435/81   Packaged device or kit

Y10S 436/80   Fluorescent dyes, e.g. rhod...

Y10S 436/81   Tube, bottle, or dipstick

Y10T 436/143333   Saccharide [e.g., DNA, etc.]

System, array and non-porous solid support comprising fixed or immobilized nucleic acids

First Claim

2 Assignments

Litigations

4 Petitions

Reexaminations

Accused Products

Abstract

333 Citations

238 Claims

Specification

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System, array and non-porous solid support comprising fixed or immobilized nucleic acids

First Claim

2 Assignments

Subscription Required

Subscription Required

Litigations

4 Petitions

Subscription Required

Reexaminations

Accused Products

Subscription Required

Abstract

333 Citations

238 Claims

Specification

Subscription Required

Use Cases

Quick Links

Others