Design and synthesis of a photocleavable biotinylated nucleotide for DNA analysis by mass spectrometry

摘要: We report here the design, synthesis and evaluation of a novel photocleavable (PC) biotinylated nucleotide analog, dUTP-PC-Biotin, for DNA polymerase extension reaction to isolate products mass spectrometry (MS) analysis. This analog has biotin moiety attached 5-position 2′-deoxyribouridine 5′-triphosphate via 2-nitrobenzyl linker. have demonstrated that dUTP-PC-Biotin can be faithfully incorporated by Thermo Sequenase into growing strand in its incorporation does not hinder addition subsequent nucleotide. Therefore, fragments generated using efficiently isolated streptavidin-coated surface recovered near-UV light irradiation at room temperature mild condition further analysis without any chemicals or heat. Single multiple primer reactions were performed generate MALDI-TOF MS Such analogs carry linker will allow isolation purification under conditions MS-based genetic sequencing multiplex single polymorphism (SNP) detection. Furthermore, these should also useful isolating DNA–protein complexes non-denaturing conditions.