Nanopore sensing is a powerful tool for the detection of biomolecules. Solid-state nanopores act as single-molecule sensors that can function in harsh conditions. Their resilient nature makes them attractive candidates for taking this technology into the field to measure environmental samples for life detection in space and water quality monitoring. Here, we discuss the fabrication of silicon nitride pores from ∼1.6 to 20 nm in diameter in 20-nm-thick silicon nitride membranes suspended on glass chips and their performance. We detect pure laboratory samples containing a single analyte including DNA, BSA, microRNA, TAT, and poly-D-lys-hydrobromide. We also measured an environmental (mixed-analyte) sample, containing Antarctic dirt provided by NASA Ames. For DNA measurements, in addition to using KCl and NaCl solutions, we used the artificial (synthetic) seawater, which is a mixture of different salts mimicking the composition of natural seawater. These samples were spiked with double-stranded DNA (dsDNA) fragments at different concentrations to establish the limits of nanopore sensitivity in candidate environment conditions. Nanopore chips were cleaned and reused for successive measurements. A stand-alone, 1-MHz-bandwidth Chimera amplifier was used to determine the DNA concentration in artificial seawater that we can detect in a practical time scale of a few minutes. We also designed and developed a new compact nanopore reader, a portable read-out device with miniaturized fluidic cells, which can obtain translocation data at bandwidths up to 100 kHz. Using this new instrument, we record translocations of 400 bp, 1000 bp, and 15000 bp dsDNA fragments and show discrimination by analysis of current amplitude and event duration histograms.
Ultrathin nanopore sensors allow single-molecule and
polymer measurements
at sub-microsecond time resolution enabled by high current signals
(∼10–30 nA). We demonstrate for the first time the experimental
probing of the ultrafast translocation and folded dynamics of double-stranded
DNA (dsDNA) through a nanopore at 10 MHz bandwidth with acquisition
of data points per 25 ns (150 MB/s). By introducing a rigorous algorithm,
we are able to accurately identify each current level present within
translocation events and elucidate the dynamic folded and unfolded
behaviors. The remarkable sensitivity of this system reveals distortions
of short-lived folded states at a lower bandwidth. This work revisits
probing of dsDNA as a model polymer and develops broadly applicable
methods. The combined improvements in sensor signals, instrumentation,
and large data analysis methods uncover biomolecular dynamics at unprecedentedly
small time scales.
The interaction between nanoparticles and zwitterionic supported lipid bilayers is a multi-step process, with specific ions exerting their influences on electrostatic-driven NP deposition and hydrophobicity-induced membrane disruption.
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