Heterocyclic FRETdye cassettes for labeling biological molecules and their use in DNA sequencing
AssigneeAmersham Biosciences AB
InventorsSatyam Nampalli, Weihong Zhang, Sudhakar Rao, Shiv Kumar
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Abstract
Exploitation of suitably functionalized heterocyclic molecules, in the design and synthesis of Fluorescence Resonance Energy Transfer (FRET) cassettes and their corresponding dideoxynucleotide terminators culminated into efficient reagents for DNA sequencing. Additionally, these FRET cassettes/terminators, of the present invention, derived from different classes of heterocyclic systems have high potential to be used for general labelling of biological molecules to generate highly sensitized signals. Their preparation, energy transfer efficiency, and use as labels, specifically, in DNA sequencing reactions is disclosed.
Claims19
An energy transfer dye labeled cassette of the formula:
- wherein
- D is a donor dye selected from the group consisting of xanthine dyes, rhodamine dyes, and cyanine dyes;
- L1 is a functional group selected from the group consisting of C1-C20alkynyl-amine, alkynol, alkenamine, alkylamine, keto, and thiol, through which D is covalently attached;
- R1, R2, R3 and R4 independently represent H, alkyl, halo, hydroxy, thio, nitro, amino or alkylamino groups;
- L2 is an amine selected from the group consisting of C1-C2alkynylamine, alkynol, alkenamine, alkylamine, keto, and thiol, through which A is covalently attached;
- X is an aldehyde, an acid, an acid chloride, an ester, hydroxymethyl, CH2O-mesylate, CH2O-triflate, mercaptomethylene, phosphoramidite or other reactive groups capable of forming a covalent bond with amine, thiol, hydroxy, or haloacetyl containing biological molecules;
- A is an acceptor dye selected from the group consisting of xanthine dyes, rhodamine dyes and cyanine dyes;
- and wherein the positions of A and D are interchangeable.
Dependencies are inferred from the claim text.