Sensitive and selective fluorometric determination of DNA by using layered hexagonal nanosheets of a covalent organic framework prepared from p-phenylenediamine and benzene-1,3,5-tricarboxaldehyde

A modified method is described for the preparation of amino-functionalized covalent organic framework nanosheets (COF-NSs). These consist of hexagonal layered sheets and were prepared from commercially available starting materials (p-phenylenediamine and benzene-1,3,5-tricarboxaldehyde). The interlayer stacking interactions between the ultra-thin COF-NSs became weak because the π stacking is destroyed by sonication. This result in the exfoliation of COF-NSs. As an application, the COF-NSs used for sensitive and selective fluorometric determination of DNA. To reach this goal, H1 and H2 hairpin-like DNA probes were chosen; H1 used Texas Red-labeled dye as a fluorescent probe. The addition of the COF-NSs, the hairpin probes was adsorbed onto the porous surface of the COFNSs. The π stacking and hydrogen-bond interactions between COFNSs and nucleic acid quench the fluorescence of the Texas red-labeled probe. The target DNA enables the recovery of the quenched fluorescence of the Texas red-labelled probe by triggering an inter-chain hybridization within hairpin probes. This results in a weaker interaction of double-stranded DNA (dsDNA) with the COFNSs. Consequently, the dsDNA detaches from the COFNSs, thereby recovering the dye’s fluorescence (excitation/emission maxima at 590/612 nm) with increasing target DNA concentration. The findings were applied to design a method for the determination of DNA that has a 2 pM detection limit. This is significantly lower than the limit of detection reported previously for 2D nanomaterial-based fluorometric DNA assays.