Phospholink nucleotides for sequencing applications
The present invention provides labeled phospholink nucleotides that can be used in place of naturally occurring nucleotide triphosphates or other analogs in template directed nucleic acid synthesis reactions and other nucleic acid reactions and various analyzes based thereon, including DNA sequencing, single base identification, hybridization assays, and others.
1. A composition, comprising a labeled phospholink nucleotide, wherein said labeled phospholink nucleotide comprises a structure that has formula:
wherein
B is a nucleobase;
S is a sugar moiety;
L is a bead comprising a fluorophore, wherein said bead is encapsulated in an organic or inorganic material;
R 1 is selected from oxygen and sulfur;
R 2 is a linker moiety; and
n is an integer selected from 0 to 9.
2. The composition of claim 1 , wherein said linker moiety is a member selected from substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocycloalkyl.
3. The composition of claim 1 , wherein said linker moiety comprises poly(ethylene glycol).
4. The composition of claim 1 , wherein said bead is a latex bead.
5. The composition of claim 1 , wherein said bead is a fluorescent bead.
6. A composition, comprising a labeled phospholink nucleotide, wherein said labeled phospholink nucleotide comprises a structure that has formula:
wherein
B is a nucleobase;
S is a sugar moiety;
L is a bead comprising a fluorophore, wherein said bead is encapsulated in a polymeric material;
R 1 is selected from oxygen and sulfur;
R 2 is a linker moiety; and
n is an integer selected from 0 to 9.
7. The composition of claim 6 , wherein said polymeric material is a member selected from polyethylene glycol and alginate.
8. A kit comprising at least four differently labeled nucleotide analogs each comprising said structure of said composition of claim 1 , a polymerase enzyme source, and instructions for using said composition with said polymerase enzyme in template directed nucleic acid synthesis.
9. The composition of claim 6 , wherein said linker moiety is a member selected from substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocycloalkyl.
10. The composition of claim 6 , wherein said linker moiety comprises poly(ethylene glycol).
11. The composition of claim 6 , wherein said bead is a latex bead.
12. The composition of claim 6 , wherein said bead is a fluorescent bead.
13. A kit comprising at least four differently labeled nucleotide analogs each comprising said structure of said composition of claim 6 , a polymerase enzyme source, and instructions for using said composition with said polymerase enzyme in template directed nucleic acid synthesis.