Isolation of genomic DNA molecule from a single cell and control its higher order structure using optical tweezers

Hidehiro Oana; Isao Hagiya; Masao Washizu; Koji Kubo; Kenichi Yoshikawa; Haruyuki Atomi; Tadayuki Imanaka

doi:10.1117/12.616498

26 August 2005 Isolation of genomic DNA molecule from a single cell and control its higher order structure using optical tweezers

Hidehiro Oana, Isao Hagiya, Masao Washizu, Koji Kubo, Kenichi Yoshikawa, Haruyuki Atomi, Tadayuki Imanaka

Author Affiliations +

Proceedings Volume 5930, Optical Trapping and Optical Micromanipulation II; 593024 (2005) https://doi.org/10.1117/12.616498
Event: Optics and Photonics 2005, 2005, San Diego, California, United States

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.

Citation Download Citation

Hidehiro Oana, Isao Hagiya, Masao Washizu, Koji Kubo, Kenichi Yoshikawa, Haruyuki Atomi, and Tadayuki Imanaka "Isolation of genomic DNA molecule from a single cell and control its higher order structure using optical tweezers", Proc. SPIE 5930, Optical Trapping and Optical Micromanipulation II, 593024 (26 August 2005); https://doi.org/10.1117/12.616498

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