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dsDNA in nanopore RNA helicase

Single Molecule Biophysics

Nanopore Force Spectroscopy (NFS)

Nanopores can be used to apply force on biomolecular complexes, such as RNA or DNA molecules or proteins. The response of the biomolecules to either stationary or time-dependent forces may reveal their structure, their interactions with other molecules and other dynamical processes. The nanopore method does not require tethering or surface immobilization of the molecules, therefore unmodified biomolecules can be studied.

 

Single Molecule Fluorescence Resonance Energy Transfer (sm-FRET)

Fluorescence resonance energy transfer is a spectroscopic method that allows probing of inter- and intra-molecular distances between a donor - acceptor fluorophore pair. During FRET an external light source (typically a laser) excites a donor molecule and the energy is transferred to the acceptor dye, which emits light at red-shifted wavelengths. Because the energy transfer efficiency strongly depends on the donor-acceptor distance, small scale (nanometer) fluctuations in the fluorophores' relative position result in large changes in the emitted light at the donor and acceptor channels. This phenomenon is widely used to obtain dynamical information on biomolecular systems, such as molecular motors and enzymes.

 

 

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