Claims
- 1. A compound of formula (I):
- 2. The compound of claim 1, wherein R is H, and L is (R1—(O)m)n—R1, in which R1 is C1-4 alkylene, C1-4 alkenylene, or C1-4 alkynylene.
- 3. The compound of claim 2, wherein R1 is CH2CH2, m is 1, and n is 1-4.
- 4. The compound of claim 3, wherein n is 1 or 2.
- 5. The compound of claim 2, wherein R1 is CH2CH2, m is 0, and n is 1-4.
- 6. A compound of formula (II):
- 7. The compound of claim 6, wherein R is H, and L is (R1—(O)m)n—R1, in which R1 is C1-4 alkylene, C1-4 alkenylene, or C1-4 alkynylene.
- 8. The compound of claim 7, wherein m is 1.
- 9. The compound of claim 8, wherein R1 is CH2CH2, and n is 1-4.
- 10. The compound of claim 9, wherein n is 1 or 2.
- 11. The compound of claim 7, wherein m is 0.
- 12. A compound of formula (III):
- 13. The compound of claim 12, wherein R is H and R′ is CH2.
- 14. The compound of claim 13, wherein B is a adenine, guanine, thymidine, uridine, or cytidine.
- 15. A method of preparing a polynucleotide, comprising:
coupling a first nucleotide or a first nucleotide oligomer to the terminal hydroxyl group of the compound of formula (II): 12 wherein L is a bond, aryl, heteroaryl, C1-14 cycloalkyl, C1-12 heterocycloalkyl, S, NH, or (R1—(O)m)n—R1, in which R1 is a C1-4 alkylene, C1-4 alkenylene, or C1-4 alkynylene, m is 1 or 0, n is 1 to 12, and SS is a first solid support, to obtain a compound of formula (IV) 13;and treating the compound of formula (IV) with a strong base to release a nucleotide or polynucleotide having a 340 amino group.
- 16. The method of claim 15, wherein the strong base is ammonia.
- 17. The method of claim 15, wherein R is H, and L is (R1—(O)m)n—R1, in which R1 is C1-4 alkylene, C1-4 alkenylene, or C1-4 alkynylene, m is 1, and n is 1 to 7.
- 18. The method of claim 15, further comprising, before the treating, sequentially coupling additional nucleotides or additional nucleotide oligomers to the first nucleotide or the first nucleotide oligomer to form an extended polynucleotide attached to the support.
- 19. The method of claim 15, further comprising covalently bonding the amino group of the released nucleotide to a second support.
- 20. The method of claim 18, wherein monomeric nucleotides are coupled.
- 21. A method to preparing a 5′-amino labeled nucleotide, comprising:
sequentially coupling nucleotides to a solid support to obtain a precursor oligonucleotide; coupling a compound of formula (I) or (III) to the precursor oligonucleotide 14 wherein R is hydrogen, C1-6 alkyl, C1-6 alkoxyl, or a halogen; and L is a bond, a nucleotide residue, aryl, heteroaryl, C1-14 cycloalkyl, C1-12 heterocycloalkyl, S, NH, or (R1—(O)m)n—R1, in which R1 is C1-4 alkylene, C1-4 alkenylene, or C1-4 alkynylene, m is 1 or 0, and n is 1 to 12, 15 wherein R is hydrogen, C1-6 alkyl, C1-6 alkoxyl, or a halogen; R′ is C1-12 alkylene, C1-12 alkenylene, or C1-12 alkynylene; and B is a nucleotide base, to the 5′ terminus of the precursor oligonucleotide thereby forming a 5′-modified oligonucleotide; and treating the 5′-modified oligonucleotide with a strong base to release the terminated oligonucleotide and a modifying group to form a 5′ amino labeled oligonucleotide.
- 22. The method of claim 21, further comprising coupling the 5′ amino labeled oligonucleotide to a substrate.
- 23. The method of claim 22, wherein the strong base is ammonia.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] Pursuant to 35 USC §119(e), this application claims the benefit of prior U.S. provisional application No. 60/325,897 filed Sep. 27, 2001.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60325897 |
Sep 2001 |
US |