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Biophys J, April 2001, p. 1932-1939, Vol. 80, No. 4
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Saint Paul, Minnesota 55108 USA
When a single molecule of double-stranded DNA is
stretched beyond its B-form contour length, the measured force shows a
highly cooperative overstretching transition. We have measured the
force at which this transition occurs as a function of temperature. To
do this, single molecules of DNA were captured between two polystyrene
beads in an optical tweezers apparatus. As the temperature of the
solution surrounding a captured molecule was increased from 11°C to
52°C in 500 mM NaCl, the overstretching transition force decreased
from 69 pN to 50 pN. This reduction is attributed to a decrease in the
stability of the DNA double helix with increasing temperature. These
results quantitatively agree with a model that asserts that DNA melting
occurs during the overstretching transition. With this model, the data
may be analyzed to obtain the change in the melting entropy
S of DNA with temperature. The observed nonlinear
temperature dependence of S is a result of the
positive change in heat capacity of DNA upon melting, which we
determine from our stretching measurements to be
Cp = 60 ± 10 cal/mol K bp, in
agreement with calorimetric measurements.
Biophys J, April 2001, p. 1932-1939, Vol. 80, No. 4
© 2001 by the Biophysical Society 0006-3495/01/04/1932/08 $2.00
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