Patent ID: 6107039
Filing Date: 2000-08-22
Classification: C07H,Y02P

Abstract:
A method of assaying a nucleic acid sequence, comprising:providing a sample comprising a nucleic acid sequence to be assayed;mixing the sample with an oligonucleotide probe thereby obtaining a treated sample, the oligonucleotide probe comprising at least one nucleotide analog of the formula: ##STR8## wherein R.sub.1 is --H, --OH, a mono, di, or triphosphate group, or --OR.sub.4 ;R.sub.2 is --H, --OH, a mono, di, or triphosphate group, a phosphoramidite group, a phosphorothioamidite group, a phosphonate group, an O-substituted monophosphate group, --OR.sub.4, or a solid support bonded via an O at the 3' position;R.sub.3 is --H, --OH, a mono, di, or triphosphate group, or --OR.sub.4 ;R.sub.4 is a lower alkyl or a protecting group; andB is a modified pyrimidine base comprising a protected thiol group attached at the 5 position on said base that is not involved in Watson-Crick base pairing or does not disrupt normal Watson-Crick base pairing, said protected thiol group being stable under conditions of chemical nucleic acid synthesis and/or conditions of enyzmatic nucleic synthesis and being convertible to a reactive thiol after said synthesis said protected thiol group selected from the group consisting of a thiodinitrophenyl group, a thioalkyldinitrophenyl group, an alkyldisulfide group, and a -S-phenylacetamidomethyl group, wherein said alkyl is a lower alkyl;exposing the treated sample to an endonuclease thereby forming fragments of the nucleic acid sequence; andidentifying fragments of the nucleic acid sequence having the oligonucleotide probe hybridized thereto.