Nanopore-based sequencers, as the fourth-generation DNA sequencing technology, have the potential to quickly and reliably sequence the entire human genome for less than $1000, and possibly for even less than $100. The single-molecule techniques used by this technology allow us to further study the interaction between DNA and protein, as well as between protein and protein. Nanopore analysis opens a new door to molecular biology investigation at the single-molecule scale. In this article, we have reviewed academic achievements in nanopore technology from the past as well as the latest advances, including both biological and solid-state nanopores, and discussed their recent and potential applications.

We propose a simple and cost-effect method, current-stimulus dielectric breakdown, to manipulate the 3D shapes of the nanochannels in 20-nm-thick SiNx membranes. Besides the precise control of nanopore size, the cone orientation can be determined by the pulse polarity. The cone angle of nanopores can be systematically tuned by simply changing the stimulus pulse waveform, allowing the gradual shape control from conical to obconical. After they are formed, the cone angle of these nanopores can be further tuned in a certain range by adjusting the widening pulse. Such size and 3D shape controllable abiotic nanopores can construct a constriction in the nanochannel and hence produce a sub-nm “sensing zone” to suit any desired bio-sensing or precise DNA sequencing. Using these conical nanopores, 20-nt ssDNA composed of homopolymers (poly(dA)20, poly(dC)20, and poly(dT)20) can be clearly differentiated by their ionic current signals.

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Antibiotics as emerging environmental contaminants, are widely used in both human and veterinary medicines. A solid-state nanopore sensing method is reported in this article to detect Tetracycline, which is based on Tet-off and Tet-on systems. rtTA (reverse tetracycline-controlled trans-activator) and TRE (Tetracycline Responsive Element) could bind each other under the action of Tetracycline to form one complex. When the complex passes through nanopores with 8 ~ 9 nanometers in diameter, we could detect the concentrations of Tet from 2 ng/mL to 2000 ng/mL. According to the Logistic model, we could define three growth zones of Tetracycline for rtTA and TRE. The slow growth zone is 0–39.5 ng/mL. The rapid growth zone is 39.5−529.7 ng/mL. The saturated zone is > 529.7 ng/mL. Compared to the previous methods, the nanopore sensor could detect and quantify these different kinds of molecule at the single-molecule level.