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We stained the λ-DNA (Gibco BRL, Gaithersburg, MD) with YOYO-1 (Molecular Probes, Eugene, OR) at a dye:base pair ratio of 1:4 for > 1 hour. The persistence length of native DNA is ∼53 nm (18). When stained, the contour length increases by up to ∼35% (to ∼22 μm for λ-DNA) [T. T. Perkins, D. E. Smith, R. G. Larson, S. Chu, Science 268, 83 (1995)]. Experiments were performed in a pH 8 buffer consisting of 10 mM tris-HCl, 2 mM EDTA, 10 mM NaCl, 4% β-mercaptoethanol, glucose oxidase (∼50 μg/ml). and catalase (∼10 μg/ml) [10 to 18% (w/w) glucose and 40 to 55% (w/w) sucrose]. The viscosity of each solution was measured with a viscometer and was adjusted by varying the sugar concentrations. The experiments were performed at room temperature (∼23°C).
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3543031114
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note
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We used a Nikon X60, 1.2 numerical aperture water immersion objective, a Zeiss 60-mm-to-infinity-corrected conversion lens, a x0.25 Zeiss tube lens, a Hamamatsu microchannel plate intensifier, and a Phillips video camera.
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55
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3543048072
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The extension data were smoothed by Gaussian weighted averages of adjacent frames
-
The extension data were smoothed by Gaussian weighted averages of adjacent frames.
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56
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3543002969
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note
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We determined the relaxation time to be 3.9 s in a 41-cP sugar solution by observing the relaxation of stretched molecules as described previously (10). The relaxation time in the sugar solutions of other viscosities was assumed to be proportional to the ratio of the viscosities.
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0000066563
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The average extension observed at zero residence time is about 2.5 μm (Fig. 2). The observed coil size appears larger than the known size of about 1.4 μm (twice the radius of gyration) because of a blooming effect in the intensified video camera when objects of saturating brightness were imaged [D. E. Smith, T. T. Perkins, S. Chu, Macromolecules 29, 1372 (1996); Phys. Rev. Lett. 75, 4146 (1995)]. The same effect was noted previously [S. B. Smith, P. K. Aldridge, J. B. Callis, Science 243, 203 (1989)] and must be kept in mind when analyzing the data.
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The average extension observed at zero residence time is about 2.5 μm (Fig. 2). The observed coil size appears larger than the known size of about 1.4 μm (twice the radius of gyration) because of a blooming effect in the intensified video camera when objects of saturating brightness were imaged [D. E. Smith, T. T. Perkins, S. Chu, Macromolecules 29, 1372 (1996); Phys. Rev. Lett. 75, 4146 (1995)]. The same effect was noted previously [S. B. Smith, P. K. Aldridge, J. B. Callis, Science 243, 203 (1989)] and must be kept in mind when analyzing the data.
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(1995)
Phys. Rev. Lett.
, vol.75
, pp. 4146
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62
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0024575851
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The average extension observed at zero residence time is about 2.5 μm (Fig. 2). The observed coil size appears larger than the known size of about 1.4 μm (twice the radius of gyration) because of a blooming effect in the intensified video camera when objects of saturating brightness were imaged [D. E. Smith, T. T. Perkins, S. Chu, Macromolecules 29, 1372 (1996); Phys. Rev. Lett. 75, 4146 (1995)]. The same effect was noted previously [S. B. Smith, P. K. Aldridge, J. B. Callis, Science 243, 203 (1989)] and must be kept in mind when analyzing the data.
-
(1989)
Science
, vol.243
, pp. 203
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Smith, S.B.1
Aldridge, P.K.2
Callis, J.B.3
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64
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3542997549
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R. Larson, in preparation
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R. Larson, in preparation.
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66
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84911793244
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See R. G. Winkler, P. Reineker, M. Schreiber, Europhys. Lett. 8, 493 (1989) for an attempt to calculate the entropic elastic forces dynamically. However, this model does not consider the dynamics in flow or allow for features such as folds.
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(1989)
Europhys. Lett.
, vol.8
, pp. 493
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Winkler, R.G.1
Reineker, P.2
Schreiber, M.3
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67
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0032488870
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E. W.-G. Diau, J. L. Herek, Z. H. Kim, A. H. Zewail, Science 279, 847 (1998).
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(1998)
Science
, vol.279
, pp. 847
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Diau, E.W.-G.1
Herek, J.L.2
Kim, Z.H.3
Zewail, A.H.4
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69
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3543009681
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note
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We acknowledge assistance from H. Babcock, W. Fann, R. Larson, T. Perkins, and W. Volkmuth. This work was supported in part by the U.S. Air Force Office of Scientific Research, National Science Foundation (NSF), the Human Frontiers Foundation, and by an endowment established by Theodore and Frances Geballe. D.E.S. was supported by a fellowship from the NSF Program in Mathematics and Molecular Biology.
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