A novel, low-cost, and disposable thread-based electrofluidic analytical method employing isotachophoresis (ITP) was developed for demonstrating surface DNA hybridization. This approach was based on graphene oxide (GO) surface-functionalized zones on nylon threads as a binding platform to trap a fluorescently labeled isotachophoretically focused single-stranded DNA (ssDNA) band, resulting in quenching of the fluorescence, which signaled quantitative trapping. In the event of an isotachophoretically focused complementary DNA (cDNA) band passing over the GO-trapped ssDNA zone, surface hybridization of the ssDNA and cDNA to form double-stranded DNA (dsDNA) band occurred, which is released from the GO-coated zones, resulting in restoration of the fluorescent signal as it exits the GO band and migrates further along the thread. This controllable process demonstrates the potential of the GO-functionalized thread-based microfluidic analytical approach for DNA hybridization and its visualization, which could be adapted into point-of-care (POC) diagnostic devices for real-world applications.
Australian Centre for Research on Separation Science (ACROSS) and ARC Centre of Excellence for Electromaterials Science (ACES), School of Natural Sciences (Chemistry), University of Tasmania, Hobart 7001, Tasmania, Australia.