Preparation and Detection of Designed DNA Carriers for Nanopore Sensing.

Single-molecule detection is a powerful strategy for early disease diagnosis, allowing the specific identification of nucleic acids and proteins. While nanopore technology enables fluorescent label-free, high-resolution analysis of individual biomolecules, its lack of intrinsic molecular selectivity limits performance in complex biological samples. The DNA carrier strategy addresses this limitation by introducing target recognition capability, incorporating molecular barcodes, and enhancing signal clarity in complex backgrounds. When combined with low-noise, high-resolution glass nanopores, these carriers enable specific and multiplexed biomarker detection for early-stage cancer and other diseases. This chapter presents a detailed and standardized protocol for preparing and detecting assembled DNA carriers using nanopores, providing an accessible workflow for broader adoption in single-molecule sensing and molecular diagnostics.