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Proceedings Paper

A general method for manipulating DNA sequences from any organism with optical tweezers
Author(s): Derek N. Fuller; Gregory J. Gemmen; John Peter Rickgauer; Aurelie DuPont; Rachel Millin; Pierre Recouvreux; Allen L. Schweitzer; Douglas E. Smith
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Paper Abstract

Here we describe and characterize a method for manipulating desired DNA sequences from any organism with optical tweezers. Molecules are produced from either genomic or cloned DNA by PCR using labeled primers and are tethered between two optically trapped microspheres. We demonstrate that human, insect, plant, bacterial, and viral sequences ranging from ~10 to 40 kbp can be manipulated. Force-extension measurements show that these constructs exhibit uniform elastic properties in accord with the expected contour lengths for the targeted sequences. Detailed protocols for preparing and manipulating these molecules are presented, and tethering efficiency is characterized as a function of DNA concentration, ionic strength, and pH. Attachment strength is characterized by measuring the unbinding time distribution as a function of applied force.

Paper Details

Date Published: 26 August 2005
PDF: 10 pages
Proc. SPIE 5930, Optical Trapping and Optical Micromanipulation II, 593013 (26 August 2005); doi: 10.1117/12.618068
Show Author Affiliations
Derek N. Fuller, Univ. of California, San Diego (United States)
Gregory J. Gemmen, Univ. of California, San Diego (United States)
John Peter Rickgauer, Univ. of California, San Diego (United States)
Aurelie DuPont, Univ. of California, San Diego (United States)
Rachel Millin, Univ. of California, San Diego (United States)
Pierre Recouvreux, Univ. of California, San Diego (United States)
Allen L. Schweitzer, Univ. of California, San Diego (United States)
Douglas E. Smith, Univ. of California, San Diego (United States)


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

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