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Proceedings Paper

Isolation of genomic DNA molecule from a single cell and control its higher order structure using optical tweezers
Author(s): Hidehiro Oana; Isao Hagiya; Masao Washizu; Koji Kubo; Kenichi Yoshikawa; Haruyuki Atomi; Tadayuki Imanaka
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Paper Abstract

In this report, we describe a noninvasive methodology for manipulating single Mb-size whole-genome DNA molecules. Cells were subjected to osmotic shock and the genome DNA released from the burst cells was transferred to a region of higher salt concentration between cover slips using optical tweezers. The transferred genome DNA exhibits a conformational transition from a compact state into an elongated state, accompanied by the change in its environment. Here, the applicability of optical tweezers to the on-site manipulation of giant genomic DNA is suggested. Next, to control the field environment more precisely, a flow chamber was made and similar investigations were carried out. In the flow chamber, the higher-order structure of individual chromosomal DNA molecules from a fission yeast that were folded by polyamine was changed to a partially unfolded form by transporting into a higher salt condition using optical tweezers. These promising methodologies demonstrated here may make it possible to recover an intact single whole-genome DNA from a cell and carry out further sequential investigations under a microscope.

Paper Details

Date Published: 27 August 2005
PDF: 8 pages
Proc. SPIE 5930, Optical Trapping and Optical Micromanipulation II, 593024 (27 August 2005); doi: 10.1117/12.616498
Show Author Affiliations
Hidehiro Oana, The Univ. of Tokyo (Japan)
Isao Hagiya, The Univ. of Tokyo (Japan)
Masao Washizu, The Univ. of Tokyo (Japan)
Koji Kubo, Kyoto Univ. (Japan)
Kenichi Yoshikawa, Kyoto Univ. (Japan)
Haruyuki Atomi, Kyoto Univ. (Japan)
Tadayuki Imanaka, Kyoto Univ. (Japan)

Published in SPIE Proceedings Vol. 5930:
Optical Trapping and Optical Micromanipulation II
Kishan Dholakia; Gabriel C. Spalding, Editor(s)

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