Fig. 1
From: Room temperature CRISPR diagnostics for low-resource settings
RPA inefficiently amplifies target below 37 °C. (a). Schematic of experiment monitoring RPA amplification efficiency using an exonuclease-sensitive probe. (b). RPA reaction progress across a range of temperatures was monitored using the exonuclease probe assay depicted in (a). Symbols represent mean (± SD) fluorescence signals of replicates (n = 3). Filled symbols indicate reactions conducted in the presence of target. Control reactions carried out in the absence of HPV-16 target (open symbols) are mostly obscured. (c) Fluorescence signals (mean ± SD) observed at 30 min from time course in (b) are displayed. (d) Schematic of experiment using qPCR on quenched RPA reactions to monitor RPA amplification efficiency. (e). RPA reaction progress at the indicated temperatures was monitored using the qPCR assay depicted in (d). Symbols represent mean (± SD) copies of replicates (n = 3). (f). RPA amplicon generation was quantified by qPCR for RPA reactions quenched after 30 min of incubation at the indicated temperatures. Reactions conducted in the absence of dNTPs reflect input target concentrations. Symbols represent mean (± SD) copies of replicates (n = 3).
