Homogeneous detection of single rolling circle replication products

摘要: We describe a simple and straightforward approach for homogeneous isothermal detection of individual rolling circle replication (RCR) products, which represent padlock probe circularization events. The RCR products constitute tens kilobases long single-stranded tandem repeated copies the sequence, in solution, they fold into micrometer-sized random coils. method is based on local enrichment fluorescence-labeled probes that hybridize to coiled compared concentration free solution. present detailed characterization using highly sensitive fast microscopy setup. At 10(4)-fold excess label, we were able detect follow at signal-to-background noise ratio 27. This high leaves room analysis device higher speeds or lower labeling ratios.