| 摘要 |
Provided are novel nucleotides, nucleoside, and their derivatives described herein, that can be used in DNA sequencing technology and other types of DNA analysis. In one embodiment, the nucleotide or nucleoside with an unprotected 3′-OH group is derivatized at the nucleobase to include a fluorescent dye attached via a linker to a photocleavable terminating group. The photocleavable-fluorescent group is designed to terminate DNA synthesis as well as be cleaved so that DNA oligomers can be sequenced efficiently in a parallel format. The design of such rapidly cleavable fluorescent groups on nucleotides and nucleosides can enhance the speed and accuracy of sequencing of large oligomers of DNA in parallel, to allow rapid whole genome sequencing, and the identification of polymorphisms and other valuable genetic information, as well as allowing further manipulation and analysis of nucleic acid molecules in their native state following cleavage of the fluorescent group. |
| 主权项 |
1. A compound according to the following formula or a salt thereof: wherein R1 is H, monophosphate, diphosphate, triphosphate, or alpha thiotriphosphate; R2 is H or OH; the Base is cytosine, uracil, thymine, adenine, guanine, or naturally occurring derivatives thereof; the Cleavable terminating moiety-Linker-Dye is a group of the formula:wherein
R3 and R4 are each independently H, a C1-C12 straight chain or branched alkyl, a C2-C12 straight chain or branched alkenyl or polyenyl, a C2-C12 straight chain or branched alkynyl or polyalkynyl, or an aromatic group, with the proviso that at least one of R3 and R4 is H; R5, R6, R7 and R8 are each independently H, OCH3, NO2, CN, a halide, a C1-C12 straight chain or branched alkyl, a C2-C12 straight chain or branched alkenyl or polyenyl, a C2-C12 straight chain or branched alkynyl or polyalkynyl, an aromatic group, or the Linker-Dye group, with the proviso that at least one of R5, R6, R7, and R8 is the Linker-Dye group; the Cleavable terminating moiety imparts polymerase termination properties to the compound and is attached to the Base by a direct bond or a linkage group; the Linker and the linkage group are, independently, a bifunctional group; and the Dye is a fluorophore. |
