A general method for manipulating DNA sequences from any organism with optical tweezers

Derek N. Fuller; Gregory J. Gemmen; John Peter Rickgauer; Aurelie DuPont; Rachel Millin; Pierre Recouvreux; Allen L. Schweitzer; Douglas E. Smith

doi:10.1117/12.618068

26 August 2005 A general method for manipulating DNA sequences from any organism with optical tweezers

Derek N. Fuller, Gregory J. Gemmen, John Peter Rickgauer, Aurelie DuPont, Rachel Millin, Pierre Recouvreux, Allen L. Schweitzer, Douglas E. Smith

Author Affiliations +

Proceedings Volume 5930, Optical Trapping and Optical Micromanipulation II; 593013 (2005) https://doi.org/10.1117/12.618068
Event: Optics and Photonics 2005, 2005, San Diego, California, United States

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.

Citation Download Citation

Derek N. Fuller, Gregory J. Gemmen, John Peter Rickgauer, Aurelie DuPont, Rachel Millin, Pierre Recouvreux, Allen L. Schweitzer, and Douglas E. Smith "A general method for manipulating DNA sequences from any organism with optical tweezers", Proc. SPIE 5930, Optical Trapping and Optical Micromanipulation II, 593013 (26 August 2005); https://doi.org/10.1117/12.618068

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