%0 Journal Article
%T A High
%A Hannah Mack
%A Mridu Sinha
%A Stephanie I. Fraley
%A Todd P. Coleman
%J SLAS TECHNOLOGY: Translating Life Sciences Innovation
%@ 2472-6311
%D 2018
%R 10.1177/2472630318769846
%X DNA melting analysis provides a rapid method for genotyping a target amplicon directly after PCR amplification. To transform melt genotyping into a broad-based profiling approach for heterogeneous samples, we previously proposed the integration of universal PCR and melt analysis with digital PCR. Here, we advanced this concept by developing a high-resolution digital melt platform with precise thermal control to accomplish reliable, high-throughput heat ramping of microfluidic chip digital PCR reactions. Using synthetic DNA oligos with defined melting temperatures, we characterized sources of melting variability and minimized run-to-run variations. Within-run comparisons throughout a 20,000-reaction chip revealed that high-melting-temperature sequences were significantly less prone to melt variation. Further optimization using bacterial 16S amplicons revealed a strong dependence of the number of melting transitions on the heating rate during curve generation. These studies show that reliable high-resolution melt curve genotyping can be achieved in digital, picoliter-scale reactions and demonstrate that rate-dependent melt signatures may be useful for enhancing automated melt genotyping
%K point-of-care testing (POCT)
%K high-throughput chemistry
%K automated biology
%K engineering
%K informatics and software
%U https://journals.sagepub.com/doi/full/10.1177/2472630318769846