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Biophys J, March 2002, p. 1537-1553, Vol. 82, No. 3
Laboratoire de Physique de la Matière Condensée, Ecole Normale Supérieure, 75005 Paris, France
Force measurements are performed on single DNA molecules
with an optical trapping interferometer that combines subpiconewton force resolution and millisecond time resolution. A molecular construction is prepared for mechanically unzipping several
thousand-basepair DNA sequences in an in vitro configuration. The force
signals corresponding to opening and closing the double helix at low
velocity are studied experimentally and are compared to calculations
assuming thermal equilibrium. We address the effect of the stiffness on the basepair sensitivity and consider fluctuations in the force signal.
With respect to earlier work performed with soft microneedles, we
obtain a very significant increase in basepair sensitivity: presently,
sequence features appearing at a scale of 10 basepairs are observed.
When measured with the optical trap the unzipping force exhibits
characteristic flips between different values at specific positions
that are determined by the base sequence. This behavior is attributed
to bistabilities in the position of the opening fork; the force flips
directly reflect transitions between different states involved in the
time-averaging of the molecular system.
Biophys J, March 2002, p. 1537-1553, Vol. 82, No. 3
© 2002 by the Biophysical Society 0006-3495/02/03/1537/17 $2.00
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