Solid-phase synthesis of new ribo and deoxyribo BrdU probes for labeling and detection of nucleic acids

A facile and efficient solid-phase synthesis of BrdU-14-UTP and BrdU-14-dUTP is reported. The method involves a nucleophilic addition of CPG-supported Br-dU with a 5′-carboxy modifier amidite, followed by the coupling with AA-UTP or AA-dUTP and finally the solid-support is cleaved under basic conditions. The newly prepared non-radioactive probes may have important applications in immunocytochemistry spatially in labeling and detection assays. BrdU moiety is coupled with nucleotides like dUTP and UTP and incorporated into nucleic acids using generally available polymerase like Taq polymerase or RNA polymerase. The probes thus generated can be used in standard blotting and hybridization procedures including in situ hybridization and detection with anti-BrdU conjugates.