Labelled nucleotides
The invention provides a nucleotide or nucleoside having a base attached to a detectable label via a cleavable linker, characterised in that the cleavable linker contains a moiety selected from the group comprising: Formula (I) (wherein X is selected from the group comprising O, S, NH and NQ wherein Q is a C 1-10 substituted or unsubstituted alkyl group, Y is selected from the group comprising O, S, NH and N(allyl), T is hydrogen or a C 1-10 substituted or unsubstituted alkyl group and * indicates where the moiety is connected to the remainder of the nucleotide or nucleoside).
1. A label-linker construct comprising the structure:
wherein R is an optionally present electron-withdrawing group; Fluor is a fluorophore; and the dotted line connecting the Fluor and the benzene ring indicates that substitution may be at any free position on the benzene ring and further atoms not shown maybe present.
2. The label-linker construct of claim 1 , wherein R is not present.
3. The label-linker construct of claim 1 , wherein R is a nitro group.
4. The labeled-linker construct of claim 1 , wherein R is a fluoro group.
5. The label-linker construction of claim 1 , comprising the structure:
6. The label-linker construct of claim 5 , wherein Fluor comprises a cyanine dye, a rhodamine dye, a coumarin dye, a fluorescein dye, an Alexa dye, a BODIPY dye, an acridine dye, a pyrene dye, or a benzanthracene dye.
7. The label-linker construct of claim 6 , wherein Fluor is a rhodamine dye.
8. The label-linker construct of claim 6 , wherein Fluor is a cyanine dye.
9. The label-linker construct of claim 8 , wherein the cyanine dye is a Cy5 dye.
10. The label-linker construct of claim 6 , wherein the Fluor is attached by reacting an N-hydroxysuccinimide ester of the Fluor with an amino moiety of the linker.
11. The label-linker construct of claim 5 , wherein the dotted line connecting the Fluor and the benzene ring comprises one or more spacer units.
12. The label-linker construct of claim 11 , wherein the spacer unit comprises —O—, —NH—, or —N(alkyl)—.
13. The label-linker construct of claim 5 , comprising the linker moiety:
14. The label-linker construct of claim 13 , comprising the structure:
15. The label-linker construct of claim 6 , wherein the label-linker is in an aqueous solution.
16. The label-linker of claim 15 , wherein the aqueous solution comprises a water soluble phosphine.
17. The label-linker of claim 16 , wherein the water soluble phosphine is a trialkyl phosphine.
18. The label-linker of claim 17 , wherein the trialkyl phosphine is derivatized with one or more hydroxyl functionalities.
19. The label-linker of claim 15 , wherein the aqueous solution further comprises a second label-linker construct, wherein the second label-linker construct comprises a different fluorophore than the Fluor.
20. The label-linker of claim 16 , wherein the aqueous solution further comprises a second label-linker construct, wherein the second label-linker construct comprises a different fluorophore than the Fluor.
21. The label-linker of claim 1 , wherein Fluor comprises a cyanine dye, a rhodamine dye, a coumarin dye, a fluorescein dye, an Alexa dye, a BODIPY dye, an acridine dye, a pyrene dye, or a benzanthracene dye.
22. The label-linker of claim 13 , wherein Fluor comprises a cyanine dye, a rhodamine dye, a coumarin dye, a fluorescein dye, an Alexa dye, a BODIPY dye, an acridine dye, a pyrene dye, or a benzanthracene dye.