Application of Padlock and Selector Probes in Molecular Medicine

Publisher Summary This chapter describes assays based on padlock and selector probes, both employing DNA ligase-assisted/DNA circularization reactions and discusses their applications. The assays are used for large-scale genotyping and amplification of targeted parts of the genome, DNA copy-number analysis, infectious diagnostics, single-molecule detection, and in situ analysis with single-nucleotide resolution. The intramolecular DNA circularization assays have important applications in molecular medicine. The targeted multiplexed genome amplification approaches have the potential to become a standard pre-preparation step for high-throughput DNA sequencers—particularly in applications where the source of DNA is limiting, such as in tumor biopsies, and where really deep sequence coverage is desired. The circularization techniques combined with RCA have unique and promising properties for rapid diagnostics and its application areas include infectious diagnostics and high-performance multiplexed quantitative expression analyses. The single-molecule in situ genotyping technique finds a broad application in biomedical research and diagnostics. Since the approach is multiplexible, it could also be applied in cell differentiation studies to determine the co-localization of several lineage-specific transcripts in developing embryos or organs. Some unique properties of circularization approaches and the advantages they bring are also discussed.