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Characterization of rodlike DNA fragments
Author(s) -
Thomas M. Record,
Woodbury Charles P.,
Inman Ross B.
Publication year - 1975
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.1975.360140212
Subject(s) - chemistry , molecular mass , sedimentation coefficient , analytical chemistry (journal) , crystallography , persistence length , sedimentation equilibrium , macromolecule , intrinsic viscosity , molecule , solvent , mass fraction , molar mass distribution , chromatography , ultracentrifuge , polymer , organic chemistry , biochemistry , enzyme
Native calf thymus DNA was sheared by sonication in a viscous solvent to the molecular‐weight range from 3 × 10 4 to 3 × 10 5 daltons, and fractionated by gel chromatography. Number and weight average molecular weights ( M̄ n and M̄ w ) were determined for individual fractions by electron microscopy; the ratio M̄ w / M̄ n for the peak fraction is approximately 1.1. Sedimentation coefficients ( s 0 20, w ) of these fractionated samples show an approximately linear dependence on the logarithm of the molecular weight M̄ w . This behavior is that expected for rodlike molecules, and is in quantitative agreement with the theory of Yamakawa and Fujii [(1973) Macromolecules 6 , 407–415] for the sedimentation coefficient of a wormlike chain with a persistence length of 625 Å, a diameter of 25 Å, and a mass per unit length of 195 daltons/Å. It appears that the wormlike coil model, without excluded volume, can represent the sedimentation behavior of DNA over the entire conformational range from rigid rod to flexible coil, using the above parameters. Equilibrium melting curves were determined for various fractions in aqueous 2.4 M tetraethylammonium bromide. A substantial broadening of the transition and decrease of the melting temperature were observed with decreasing molecular weight. Empirical expressions have been obtained relating both the transition temperature and breadth in this solvent to molecular weight.