A Lab-on-a-Chip for the Extraction and Analysis of Single Molecules of DNA from Biological Media

DNA extraction is a critical prerequisite for reliable downstream analyses such as Polymerase Chain Reaction (PCR), sequencing, and genotyping. Conventional methods often require labor-intensive protocols, large sample volumes, or costly automation. Microfluidic approaches offer an alternative by reducing reagent consumption and enabling faster, more integrated workflows. Here, we present a passive lab-on-a-chip device that performs DNA extraction from complex biological media and enables subsequent on-chip single-molecule analysis. The chip integrates a magnetophoresis-based solid-phase extraction module with a fluorescence detection section capable of quantifying DNA molecules in microchannels and visualizing stretched molecules in nanochannels. The multi-level micro/nanofluidic architecture is fabricated in polymer using a single-step nanoimprinting process with a total manufacturing time of two minutes per chip, enabling scalable production. As a proof of concept, the device extracted DNA from samples spiked into buffer or plasma. On-chip transfer efficiency of DNA–bead complexes to the elution buffer reached 86%, and quantitative analysis of the recovered liquid showed an overall extraction efficiency of 40% (including DNA recovery off-chip), with intact 48 kbp DNA confirmed in both micro- and nanochannel measurements. This platform offers a promising foundation for point-of-care and point-of-interest applications, where integrated DNA extraction and analysis can reduce sample loss and support streamlined, automated workflows.