What is claimed is:
1. A carbon film comprising an acicular carbon layer and a filamentous carbon layer which is covered on the acicular carbon layer.
2. A carbon film as claimed in claim 1, wherein the acicular carbon layer comprises a plurality of conical andor pyramidal shape projections.
3. A carbon film as claimed in claim 2, wherein the projections have bottom widths that are not greater than 200 nm.
4. A carbon film as claimed in claim 1, wherein the filamentous carbon layer is formed by at least one carbon filament that is not greater than 10 nm in filament width.
5. A film for use in emitting electrons, the film comprising a filamentous layer formed by a plurality of filaments.
6. A film as claimed in claim 5, wherein the filamentous layer is attached to an acicular layer.
7. A film as claimed in claim 6, wherein the acicular carbon layer comprises a plurality of acicular projections.
8. A film as claimed in claim 7, wherein the acicular projections have conical andor pyramidal shapes.
9. A film as claimed in claim 5, wherein the filamentous layer is composed of carbon.
10. A film as claimed in claim 6, wherein both the filamentous layer and the acicular layer are formed by carbon.
11. A field emission cathode comprising an emitter for emitting electrons, wherein the emitter comprises a carbon film which has a composite layer structure.
12. A field emission cathode as claimed in claim 11, wherein the composite layer structure has an acicular carbon layer and a filamentous carbon layer which is covered on the acicular carbon layer.
13. A field emission cathode as claimed in claim 12, wherein the acicular carbon layer comprises a plurality of conical andor pyramidal shape projections.
14. A field emission cathode as claimed in claim 13, wherein the filamentous carbon layer is formed by at least one carbon filament that is not greater than 10 nm in filament width.
15. A method of manufacturing a composite carbon film, comprising the steps of:
depositing a carbon film;
forming an acicular carbon layer composed of acicular projections on a surface of the carbon film; and
changing the acicular carbon layer into a filamentous carbon layer comprising at least one filament to form the composite carbon film.
16. A method as claimed in claim 15, wherein the forming step comprising the step of:
carrying out plasma processing of the carbon film within a hydrogen gas to form the acicular carbon layer;
the changing step comprising the step of:
carrying out plasma processing the acicular carbon layer within a hydrogen gas to form the filamentous carbon layer attached to the acicular carbon layer and to thereby form the composite carbon film.
17. A method as claimed in claim 15, wherein the forming step comprising the step of:
carrying out plasma processing of the carbon film within a hydrogen gas and an oxygen gas to form the acicular carbon layer;
the changing step comprising the step of:
carrying out plasma processing the acicular carbon layer within a hydrogen gas to form the filamentous carbon layer attached to the acicular carbon layer and to thereby form the composite carbon film.
18. A method of manufacturing a field emission cathode which has an emitter, comprising the steps of:
depositing a carbon film;
forming an acicular carbon layer composed of acicular projections on a surface of the carbon film; and
changing the acicular carbon layer into a filamentous carbon layer comprising at least one filament to form the emitter.
19. A method as claimed in claim 18, wherein the forming step comprising the step of:
carrying out plasma processing of the carbon film within a hydrogen gas to form the acicular carbon layer;
the changing step comprising the step of:
carrying out plasma processing the acicular carbon layer within a hydrogen gas to form the filamentous carbon layer attached to the acicular carbon layer and to thereby form the emitter.
20. A method as claimed in claim 18, wherein the forming step comprising the step of:
carrying out plasma processing of the carbon film within a hydrogen gas and an oxygen gas to form the acicular carbon layer;
the changing step comprising the step of:
carrying out plasma processing the acicular carbon layer within a hydrogen gas to form the filamentous carbon layer attached to the acicular carbon layer and to thereby form the emitter.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
What is claimed is:
1. A method of producing an immobilized oligonucleotide on a substrate comprising:
covalently attaching a first nucleotide or an oligonucleotide via a C-5 oxygen of the nucleotide to the substrate.
2. The method of claim 1, wherein the first nucleotide has the formula
9
wherein X is selected from the group consisting of a photolabile protecting group and a chemically labile protecting group; R is selected from the group consisting of hydrogen, hydroxyl, and modified hydroxyl; P is selected from the group consisting of hydrogen, a phosphorous activating group, and a phosphate or derivative thereof; and Base is selected from the group consisting of pyrimidine, purine, and derivatives thereof.
3. The method of claim 2 further including removing the photolabile protecting group or the chemically labile protecting group from the C-3 oxygen of the first nucleotide to form a hydroxyl group.
4. The method of claim 3 further including covalently attaching a second nucleotide monomer or oligonucleotide via a C-5 oxygen of a nucleotide to the first nucleotide.
5. The method of claim 4 further including activating the C-3 hydroxyl group of the first nucleotide with a phosphorous activating group.
6. The method of claim 5, wherein the phosphorous activating group is chloro-N,N-diisopropylamine–cyanoethoxyphosphine or bis-N,N-diisopropylamine–cyanoethoxyphosphine
7. The method of claim 1 further including activating a terminal end of a linking group bound to the substrate.
8. The method of claim 7, wherein the first nucleotide covalently attaches to the activated terminal end of the linking group.
9. The method of claim 8, wherein the linking group has the formula (OCH2CH2)nO and n has value between about 3 and about 30.
10. The method of claim 2, wherein the base is selected from the group consisting of adenyl, guanyl, cytidyl, thymidyl, inosyl, and uridyl.
11. The method of claim 10, wherein the base is inosyl.
12. The method of claim 2, wherein X is a photolabile protecting group selected from the group consisting of NVOC, MBNPEOC, and MeNPOC.
13. The method of claim 12, wherein X is MBNPEOC.
14. The method of claim 2, wherein the first nucleotide has the formula
10
wherein R1, R2, R3 each independently is selected from the group consisting of hydrogen, C1-C10 alkyl, C2-C10 alkenyl, aryl, benzyl, and C1-C10 alkoxyl; R4 is selected from the group consisting of C1-C10 alkyl, C2-C10 alkenyl, aryl, and benzyl; and P is selected from the group consisting of hydrogen and (i-Pr)2NPOCH2CH2CN.
15. The method of claim 1, wherein the substrate is selected from the group consisting of glass, surface-modified glass, particles, and shaped gel.
16. A method of synthesizing an oligonucleotide on a substrate, comprising:
(a) contacting a modified nucleotide via a C-5 oxygen to an activated immobilized hydroxyl group to produce a covalently attached nucleotide, wherein the modified nucleotide includes a C-3 photolabile protecting group and a C-5 hydroxyl group, and wherein the activated immobilized hydroxyl group is activated with a phosphorous activating group;
(b) irradiating the covalently attached nucleotide to remove the C-3 photolabile protecting group and form a C-3 hydroxyl group;
(c) contacting the C-3 hydroxyl group of the covalently attached nucleotide with a phosphorous activating group to produce an activated immobilized hydroxyl group at the C-3 position of the covalently attached nucleotide; and
(d) repeating steps (a) to (c).
17. The method of claim 16, wherein the modified nucleotides have the formula
11
wherein X is a photolabile protecting group; R is selected from the group consisting of hydrogen, hydroxyl, and modified hydroxyl; P is hydrogen; and Base is selected from the group consisting of pyrimidine, purine, and derivatives thereof.
18. The method of claim 16, wherein the phosphorous activating group is chloro-N,N-diisopropylamine–cyanoethoxyphosphine or bis-N,N-diisopropylamine–cyanoethoxyphosphine.
19. The method of claim 16, wherein the surface hydroxyl group is the terminus of a linking group having the formula (OCH2CH2)nOH and n has value between about 3 and about 30.
20. The method of claim 16, wherein the base is selected from the group consisting of adenyl, guanyl, cytidyl, thymidyl, inosyl, and uridyl.
21. The method of claim 16, wherein the photolabile protecting group is selected from the group consisting of NVOC, MBNPEOC, and MeNPOC.
22. The method of claim 21, wherein the photolabile protecting group is MBNPEOC.
23. The method of claim 16, wherein modified nucleotides have the formula
12
wherein R1, R2, R3 each independently is selected from the group consisting of hydrogen, C1-C10 alkyl, C2-C10 alkenyl, aryl, benzyl, and C1-C10 alkoxyl; R4 is selected from the group consisting of C1-C10 alkyl, C2-C10 alkenyl, aryl, and benzyl; and P is selected from the group consisting of hydrogen and (i-Pr)2NPOCH2CH2CN.
24. A method of claim 16 further including contacting the synthesized oligonucleotide with a polymerase, wherein the polymerase is selected from a group consisting of DNA polymerases and RNA polymerases.
25. A method of claim 16, wherein the substrate is selected from the group consisting of glass, surface-modified glass, particles, and shaped gel.
26. A method of synthesizing an oligonucleotide, comprising:
(a) providing a nucleotide or an oligonucleotide having a free terminal C-3 hydroxyl and a terminal C-5 that is blocked by a group, wherein the free terminal C-3 hydroxyl is activated with a phosphorous activating group;
(b) covalently coupling a modified nucleotide via a C-5 oxygen to the activated hydroxyl group, wherein the modified nucleotide includes a C-3 photolabile protected group and a C-5 hydroxyl group;
(c) irradiating the covalently attached nucleotide to remove the C-3 photolabile protecting group and form a C-3 hydroxyl group;
(d) contacting the C-3 hydroxyl group of the covalently attached nucleotide with a phosphorous activating group to produce an activated hydroxyl group at the C-3 position;
(e) covalently coupling a modified nucleotide via a C-5 oxygen to the activated hydroxyl group of a previously covalently attached nucleotide; and
(f) repeating steps (c) to (e).
27. The method of claim 26, wherein the modified nucleotides have the formula
13
wherein X is a photolabile protecting group; R is selected from the group consisting of hydrogen, hydroxyl, and modified hydroxyl; P is hydrogen; and Base is selected from the group consisting of pyrimidine, purine, and derivatives thereof.
28. The method of claim 27, wherein the base is selected from the group consisting of adenyl, guanyl, cytidyl, thymidyl, inosyl, and uridyl.
29. The method of claim 27, wherein the photolabile protecting group is selected from the group consisting of NVOC, MBNPEOC, and MeNPOC.
30. The method of claim 27, wherein the photolabile protecting group is MBNPEOC.
31. The method of claim 27, wherein modified nucleotides have the formula
14
wherein R1, R2, R3 each independently is selected from the group consisting of hydrogen, C1-C10 alkyl, C2-C10 alkenyl, aryl, benzyl, and C1-C10 alkoxyl; R4 is selected from the group consisting of C1-C10 alkyl, C2-C10 alkenyl, aryl, and benzyl; and P is selected from the group consisting of hydrogen and (i-Pr)2NPOCH2CH2CN.
32. An oligonucleotide array, comprising:
A substrate having a plurality of addressable sites; each of the sites of the plurality having an oligonucleotide covalently attached to the substrate via its C-5 oxygen atom, wherein each site of the plurality is directly adjacent to at least one other site, and wherein the sequence of each oligonucleotide is unique among the plurality.
33. The oligonucleotide array of claim 32, wherein the substrate is selected from the group consisting of glass, surface-modified glass, and shaped gel.
34. The oligonucleotide array of claim 32, wherein the attached oligonucleotides have a density of at least 20 moleculescm2.
35. The oligonucleotide array of claim 32, wherein the attached oligonucleotides have a density of at least 50,000 moleculescm2.
36. The oligonucleotide array of claim 32, wherein the attached oligonucleotides have a density of at least 1,000,000 moleculescm2.
37. The oligonucleotide array of claim 32, wherein the unique sequence has a length of 5 bases to 40 bases.
38. A compound of the formula
15
wherein X is selected from the group consisting of a photolabile protecting group and a chemically labile protecting group; R is selected from the group consisting of hydrogen, hydroxyl, and alkoxyl; P is selected from the group consisting of hydrogen, a phosphorous activating group, a nucleotide monomer or oligomer, and a phosphate or derivative thereof; and Base is selected from the group consisting of pyrimidine, purine, and derivatives thereof.
39. The compound of claim 38 wherein the base is selected from the group consisting of adenyl, guanyl, cytidyl, thymidyl, inosyl, and uridyl.
40. The compound of claim 39, wherein the base is inosyl.
41. The compound of claim 38, wherein X a photolabile protecting group selected from the group consisting of NVOC, MBNPEOC, and MeNPOC.
42. The compound of claim 41, wherein X is MBNPEOC.
43. The compound of claim 38, wherein P is hydrogen.
44. The compound of claim 38, wherein P is a phosphate moiety bound to a linking group attached to a support surface.
45. The compound of claim 38, wherein the linking group has the formula (OCH2CH2)nO and n has value between about 3 and about 30.
46. A compound of the formula
16
wherein R1, R2, R3 each independently is selected from the group consisting of hydrogen, C1-C10 alkyl, C2-C10 alkenyl, aryl, benzyl, and C1-C10 alkoxyl; R4 is selected from the group consisting of C1-C10 alkyl, C2-C10 alkenyl, aryl, and benzyl; and P is selected from the group consisting of hydrogen and (i-Pr)2NPOCH2CH2CN.
47. The compound of claim 46, wherein P is a hydrogen atom.
48. The compound of claim 46, wherein P is phosphoramidite.
49. The compound of claim 46, wherein the base is selected from the group consisting of adenyl, guanyl, cytidyl, thymidyl, inosyl, and uridyl.
50. The compound of claim 49, wherein the base is inosyl.
51. The compound of claim 46, wherein P is an oligonucleotide.
52. A compound of the formula
17
where Y is a C-3 oxygen of a nucleic acid and R1 is methyl.