Claims
- 1. A photoreactive phosphoramidite, wherein said photoreactive phosphoramidite incorporates a first photoreactive site that undergoes 2+2 cycloaddition with a second photoreactive site when irradiated with light of an appropriate wavelength.
- 2. A photoreactive phosphoramidite having the structure
- 3. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 4. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 5. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 6. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 7. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 8. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 9. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 10. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 11. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 12. The photoreactive phosphoramidite of claim 2, wherein E is alkyl
- 13. The photoreactive phosphoramidite of claim 2, wherein R1 is —CH2CH2CN and R2 is isopropyl.
- 14. A photoreactive phosphoramidite having the structure
- 15. The photoreactive phosphoramidite of claim 14, wherein A is hexyl.
- 16. The photoreactive phosphoramidite of claim 14, wherein A is cyclohexyl.
- 17. The photoreactive phosphoramidite of claim 14, wherein A is
- 18. The photoreactive phosphoramidite of claim 14, wherein A is
- 19. The photoreactive phosphoramidite of claim 14, wherein R1 is −CH2CH2CN and R2 is isopropyl.
- 20. A photoreactive phosphoramidite having the structure
- 21. The photoreactive phosphoramidite of claim 20, wherein R1 is —CH2CH2CN and R2 is isopropyl.
- 22. A probe for use in a microarray analysis comprising a photoreactive site and an oligonucleotide, wherein said photoreactive site is incorporated into said oligonucleotide by a photoreactive phosphoramidite.
- 23. The probe of claim 22, wherein the microarray analysis is an expression or a SNP analysis.
- 24. The probe of claim 22, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 14.
- 25. The probe of claim 22, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 2.
- 26. The probe of claim 22, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 20.
- 27. A composition comprising an oligonucleotide covalently attached to a hydrogel, wherein said attachment is by a 2+2 cycloaddition between a first photoreactive site incorporated into said oligonucleotide by a photoreactive phosphoramidite and a second photoreactive site present on said hydrogel.
- 28. The composition of claim 27, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 14.
- 29. The composition of claim 27, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 2.
- 30. The composition of claim 27, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 20.
- 31. A method of making a photoreactive oligonucleotide probe comprising:
(a) a stepwise addition of nucleoside phosphoramidites to a synthesis support to form an oligonucleotide; (b) chemically coupling a photoreactive phosphoramidite with the oligonucleotide to form the photoreactive oligonucleotide probe, wherein said photoreactive oligonucleotide probe incorporates a first photoreactive site capable of undergoing 2+2 cycloaddition; (c) removing the photoreactive oligonucleotide probe from the synthesis support; and (d) optionally deprotecting and purifying said photoreactive oligonucleotide probe.
- 32. The method of claim 31, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 14.
- 33. The method of claim 31, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 2.
- 34. The method of claim 31, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 20.
- 35. A method of attaching the photoreactive oligonucleotide probe of claim (c) to a hydrogel comprising:
(a) providing the photoreactive oligonucleotide probe; (b) providing a hydrogel with a second photoreactive site; (c) covalently bonding said photoreactive oligonucleotide probe to said hydrogel by combining said photoreactive oligonucleotide probe with said hydrogel and irradiating with ultraviolet light, wherein a 2+2 cycloaddition occurs between the first and second photoreactive sites.
- 36. The method of claim 35, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 14.
- 37. The method of claim 35, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 2.
- 38. The method of claim 35, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 20.
- 39. In a method for synthesizing an oligonucleotide or nucleic acid probe for attachment to a hydrogel, the improvement comprising incorporating a photoreactive site into a phosphoramidite coupler to form a photoreactive phosphoramidite and reacting said photoreactive phosphoramidite with said oligonucleotide or nucleic acid.
- 40. The method of claim 39, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 14.
- 41. The method of claim 39, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 2.
- 42. The method of claim 39, wherein said photoreactive phosphoramidite is the photoreactive phosphoramidite of claim 20.
REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of U.S. Nonprovisional Application No. 09/928,250, filed Aug. 9, 2001, entitled “The Use and Evaluation of 2+2 Photoaddition in Immobilization of Oligonucleotides on A Three Dimensional Hydrogel Matrix,” (incorporated by reference) which is a continuation-in-part of U.S. Nonprovisional Application No. 09/344,620, filed Jun. 25, 1999, entitled “Methods and Compositions for Attachment of Biomolecules to Solid Supports, Hydrogels, and Hydrogel Arrays” (incorporated by reference).
Continuation in Parts (2)
|
Number |
Date |
Country |
Parent |
09928250 |
Aug 2001 |
US |
Child |
10185279 |
Jun 2002 |
US |
Parent |
09344620 |
Jun 1999 |
US |
Child |
09928250 |
Aug 2001 |
US |