id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-009318-zt1o1bcz	Rolando, Justin C	Real-time kinetics and high-resolution melt curves in single-molecule digital LAMP to differentiate and study specific and non-specific amplification	2020-04-17		.txt	text/plain	13716	626	45	We then develop a real-time digital LAMP (dLAMP) with high-resolution melting temperature (HRM) analysis and use this single-molecule approach to analyze approximately 1.2 million amplification events. By differentiating true and false positives, HRM enables determination of the optimal assay and analysis parameters that leads to the lowest limit of detection (LOD) in a digital isothermal amplification assay. To test this hypothesis, we used a dLAMP assay with CT DNA as the target (combined with sequencing to identify the products of bulk reactions) to analyze both specific and non-specific amplification under conditions that include clinically relevant concentrations of background human DNA. Single digital partition counts were observed at low-T m non-specific amplification in both the presence of template and the NTC and independent of hgDNA concentration ( Figure 9B and C). When using Bst 3.0 (Supplementary Figure S11F) and HRM to remove non-specific amplification, LOD tracks with the number of true-positive events.	./cache/cord-009318-zt1o1bcz.txt	./txt/cord-009318-zt1o1bcz.txt