MODIFIED NUCLEOTIDES FOR POLYNUCLEOTIDE SEQUENCING

摘要

The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3'-OH blocking group covalently attached thereto, such that the 3' carbon atom has attached a group of the structure
€ƒ€ƒ€ƒ€ƒ€ƒ€ƒ€ƒ€ƒ-O-Z
wherein Z is any of -C(R') 2 -O-R", -C(R') 2 -N(R") 2 ,-C(R') 2 -N(H)R", -C(R') 2 -S-R" and -C(R') 2 -F,
wherein each R" is or is part of a removable protecting group;
each R' is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R') 2 represents an alkylidene group of formula =C(R"') 2 wherein each R''' may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and
wherein said molecule may be reacted to yield an intermediate in which each R" is exchanged for H or, where Z is -C(R') 2 -F, the F is exchanged for OH, SH or NH 2 , preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3'OH;
with the proviso that where Z is -C(R') 2 -S-R", both R' groups are not H.