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Proceedings Paper

Pulsed-field electrophoresis of nucleic acids: ultrafast separations in ultrashort capillaries
Author(s): Michael D. Morris; Yongseong Kim; Richard W. Hammond
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Paper Abstract

Nucleic acid electrophoretic separations in unentangled water-soluble polymers are rapid and applicable over the length range 500 bp to greater than 1.5 Mbp, depending upon the polymer matrix chosen. We have been able to achieve high resolution separations in 3 - 4 minutes, even in the Mbp size range. Pulsed field electrophoresis is necessary for chains longer than about 20 kbp and is beneficial above about 2 - 3 kbp. The separation speed is a consequence of several factors: (1) the low viscosity of an unentangled polymer solution. (2) the high electric fields usable in pulsed field electrophoresis. And (3) the use of short (ca. 10 cm) capillaries. Electrophoretic measurements are guided by video fluorescence microscopy, which is used to study the dynamics of the separation process. Analysis of image sequences demonstrates that the segmental motions of DNA, which generate the size-dependent mobilities, are quite similar to those previously observed in entangled linear polymers and cross-linked gels. We describe our most recent separations and the current state of understanding of DNA dynamics in unentangled polymer matrices.

Paper Details

Date Published: 1 April 1996
PDF: 6 pages
Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); doi: 10.1117/12.237609
Show Author Affiliations
Michael D. Morris, Univ. of Michigan (United States)
Yongseong Kim, Univ. of Michigan (United States)
Richard W. Hammond, Univ. of Michigan (United States)

Published in SPIE Proceedings Vol. 2680:
Ultrasensitive Biochemical Diagnostics
Gerald E. Cohn; Steven A. Soper; C. H. Winston Chen, Editor(s)

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