COMPOSITIONS, PROBES AND CONJUGATES AND USES THEREOF
First Claim
1. A composition comprising a conjugate, said conjugate comprising a nucleotide and an ethynyl group functioning to emit a detectable signal when reacted with a compound comprising an azide group.
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Abstract
The present invention relates to compositions useful as probes and in other applications and methods of their use. In some embodiments, nucleotides are prepared and functionalized with dyes. In some embodiments a first molecule is functionalized with an alkynyl group, a second molecule is functionalized with an azide group, and said first and second molecules are mixed under conditions to form a conjugate with a 1,2,3-triazol group. In further embodiments, a nucleotide is functionalized with an alkynyl group, a dye is functionalized with an azide group, and mixing the nucleotide and the dye forms a conjugate capable of emitting light.
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Citations
16 Claims
- 1. A composition comprising a conjugate, said conjugate comprising a nucleotide and an ethynyl group functioning to emit a detectable signal when reacted with a compound comprising an azide group.
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4. A composition comprising a substituted or unsubstituted unreacted compound functioning to react with an azide to form a reacted compound with a 1,2,3-triazol group, said unreacted compound having the following formula:
- 5. A composition comprising a compound with a 1,2,3-triazol group, wherein said compound has the following formula:
- 7. A composition comprising an unreacted compound functioning to react with an azide to form a reacted compound comprising a 1,2,3-triazol group, said unreacted compound having the following formula:
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9. A method for making a nucleic acid molecule comprising a label, comprising:
- reacting a first unreacted compound, wherein said first unreacted compound is linked to a nucleic acid molecule and comprises an ethynyl group, with a second unreacted compound, wherein said second unreacted compound comprises an azide group, to create a nucleic acid molecule comprising a label.
- View Dependent Claims (10, 11, 12)
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13. A method of making 5-(4-ethynylphenyl)amino-5-oxo-pentanoic acid comprising:
- mixing 4-ethynylaniline with glutaric anhydride under conditions such that 5-(4-ethynylphenyl)amino-5-oxo-pentanoic acid is formed.
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14. A method of making N′
- -(6-aminohexyl 2-cyanoethyl diisopropylamidophosphite)-N-(4-ethynylphenyl)-pentandiamide comprising mixing N′
-(6-hydroxyhexyl)-N-(4-ethynylphenyl)-pentandiamide with 2-Cyanoethyl-N,N,N′
,N′
-tetraisopropylphosphoramidite under conditions such that N′
-(6-aminohexyl 2-cyanoethyl diisopropylamidophosphite)-N-(4-ethynylphenyl)-pentandiamide is formed.
- -(6-aminohexyl 2-cyanoethyl diisopropylamidophosphite)-N-(4-ethynylphenyl)-pentandiamide comprising mixing N′
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15. A method of making N′
- -(6-hydroxyhexyl)-N-(4-ethynylphenyl)-pentandiamide comprising;
i) mixing 5-(4-ethynylphenyl)amino-5-oxo-pentanoic acid with N-hydroxysuccinimide under conditions such that (2,5-dioxopyrrolidin-1-yl) 5-(4-ethynylphenyl)amino-5-oxo-pentanate is formed and ii) mixing (2,5-dioxopyrrolidin-1-yl) 5-(4-ethynylphenyl)amino-5-oxo-pentanate with 1-amino-6-hexanol under conditions such that N′
-(6-hydroxyhexyl)-N-(4-ethynylphenyl)-pentandiamide is formed.
- -(6-hydroxyhexyl)-N-(4-ethynylphenyl)-pentandiamide comprising;
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16. A method of making N′
- -(6-hydroxyhexyl)-N-(4-ethynylphenyl)-pentandiamide comprises i) mixing 5-(4-ethynylphenyl)amino-5-oxo-pentanoic acid with a leaving group reagent under conditions such that a compound comprising a leaving group is formed and ii) mixing said compound comprising said leaving group with 1-amino-6-hexanol under conditions such that N′
-(6-hydroxyhexyl)-N-(4-ethynylphenyl)-pentandiamide is formed.
- -(6-hydroxyhexyl)-N-(4-ethynylphenyl)-pentandiamide comprises i) mixing 5-(4-ethynylphenyl)amino-5-oxo-pentanoic acid with a leaving group reagent under conditions such that a compound comprising a leaving group is formed and ii) mixing said compound comprising said leaving group with 1-amino-6-hexanol under conditions such that N′
Specification