Patent ID: 7282581

Claim:
A labeling reactant of formula (I), useful for labeling an oligonucleotide using solid phase chemistry, wherein R is a protecting group or not present; A is either a phosphorylating moiety where L is O, S, or not present L′ is H, XCH 2 CH 2 CN or XAr, where Ar is phenyl or its substituted derivative, where the substituent is nitro or chlorine, and X is O or S; L″ is O − , S − , Cl, N(i-Pr) 2 ; or A is a solid support tethered to Z via a linker arm E″; Z is a bridge point and is formed from E is a linker arm between R and Z, E′ is a linker arm between Z and Z′, E″ is a linker arm between Z and A and E′″ is a linker arm between Z′ and G, same or different, and is formed of one to ten moieties, each moiety being selected from the group consisting of phenylene, alkylene containing 1–12 carbon atoms, ethynediyl, ether, thioether, amide, carbonyl, ester, disulfide, diaza, and tertiary amine; Z′ is a pyrimidine base selected from the group consisting of cytosine, thymine, uracil, where E′ is attached to N1 of pyrimidines, and E′″ is attached to N3 or C5 of uracil, N3 of thymine, C5 or N 4 of cytosine, and where the exocyclic functional groups of said base are protected, or Z′ is selected from the group consisting of imidazole, pyrazolo[3,4-d]pyrimidine, 4-amino-pyrazolo[3,4-d]pyrimidine, 1,2,4-triazine-3,5-dione, 5-amino-1,2,4-triazine-3-one, where E′ is attached to N1 of imidazole, N2 of 1,2,4-triazine-3,5-dione and 5-amino-1,2,4-triazine-3-one, and N7 of 4-amino-pyrazolo[3,4-d]pyrimidine and pyrazolo[3,4-d]pyrimidine, and E″ is attached to C4 or C5 of imidazole, C2 or C9 of 4-amino-pyrazolo[3,4-d]pyrimidine, C2, C4 or C9 of pyrazolo[3,4-d]pyrimidine, N4, C5 or C6 of 1,2,4-triazine-3,5-dione and N 5 or C6 of N 5 -amino 1,2,4-triazine-3,5-dione and where the exocyclic functional groups of said base are protected; G is a protected bivalent aromatic structure, tethered to two iminodiacetic acid ester groups N(CH 2 COOR″) 2 where R″ is an alkyl of 1 to 4 carbon atoms, allyl, ethyltrimethylsilyl, phenyl or benzyl, which phenyl or benzyl is substituted or unsubstituted, and one of the hydrogen atoms is substituted with E′″, and said bivalent aromatic structure is capable of absorbing light or energy and transferring the excitation energy to a lanthanide ion after said labeling reactant has been deprotected and converted to a lanthanide chelate, or G is a structure selected from the group consisting of where R″ is an alkyl of 1 to 4 carbon atoms, allyl, ethyltrimethylsilyl, phenyl or benzyl, which phenyl or benzyl is substituted or unsubstituted, and one of the hydrogen atoms is substituted with E′″, or G is a protected functional group, where the functional group is amino, aminooxy, carboxyl, thiol, and the protecting group is pthaloyl, trityl, 2-(4-nitrophenylsulfonyl)ethoxycarbonyl, fluorenylmethyloxycarbonyl, benzyloxycarbonyl, t-butoxycarbonyl or trifluoroacetuyl for amino and aminooxy, alkyl for carbonyl and alkyl or trityl for thiol, or G is a protected or unprotected organic dye, hapten or a spin label.