We present the use of an alternating current (AC) signal as a means to monitor the conductance of an α-hemolysin (αHL) pore as a DNA hairpin with a polydeoxyadenosine tail is driven into and released from the pore. Specifically, a 12 base pair DNA hairpin attached to a 50-nucleotide poly-A tail (HP-A50) is threaded into an αHL channel using a DC driving voltage. Once the HP-A50 molecule is trapped within the αHL channel, the DC driving voltage is turned off and the conductance of the channel is monitored using an AC voltage. The escape time, defined as the time it takes the HP-A50 molecule to transport out of the αHL channel, is then measured. This escape time has been monitored as a function of AC amplitude (20 to 250 mVac), AC frequency (60–200 kHz), DC drive voltage (0 to 100 mVdc), and temperature (−10 to 20 °C), in order to determine their effect on the predominantly diffusive motion of the DNA through the nanopore. The applied AC voltage used to monitor the conductance of the nanopore has been found to play a significant role in the DNAanopore interaction. The experimental results are described by a one-dimensional asymmetric periodic potential model that includes the influence of the AC voltage. An activation enthalpy barrier of 1.74 × 10−19 J and a periodic potential asymmetry parameter of 0.575 are obtained for the diffusion at zero electrical bias of a single nucleotide through αHL.
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