The project concerns optical single molecule sequencing on a highly packed array of molecules in which individual molecules are not resolvable by standard diffraction limited imaging. The sequence of molecules is obtained by super-resolving signal from individual molecules during the course of template-directed DNA synthesis. The project will: (1) synthesize and test the basic building blocks of the chemistry in order to demonstrate multi- contiguous base incorporation; (2) capture target molecules in an ordered nanoarray; (3) Super-resolve the signals within the nanoarray and increase confidence in base-calling by repetitive signal acquisition. Overall the method will reduce the cost of sequencing 10x by reducing the amount of reagents consumed and increasing throughput achievable within the chip. The method will also increase the sensitivity and accuracy of sequencing by making multiple measurements at the single molecule level, and jettisoning the parts of current sequencing workflows that introduce error.
Public Health Relevance Statement: Narrative The research proposes an innovative means to increase the cost, accuracy and sensitivity of sequencing so that it can fulfill the needs of sequencing in healthcare. This should impact public health by enabling better genetic epidemiological studies and could impact individuals by facilitating the personalization of their healthcare.
Project Terms: Adoption; analog; base; Binding; Chemistry; Clinic; cost; Data Quality; density; deoxyribonucleoside triphosphate; DNA; DNA biosynthesis; Future; genetic epidemiology; Healthcare; Image; Individual; innovation; Label; Length; Measurement; Methods; Nanoarray Analytical Device; nanoscale; novel; Nucleotides; Oligonucleotides; Optics; Outcome; Penetration; Phase; phase 1 study; Public Health; Reaction; Reagent; Research; Resolution; scaffold; Signal Transduction; single molecule; Speed; Surface; Technology; Testing
