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Biophys. J. BioFAST: First Published September 8, 2006. doi:10.1529/biophysj.106.089524
© 2006 by the Biophysical Society.

A more recent version of this article appeared on December 1, 2006.
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SPECTROSCOPY, IMAGING, OTHER TECHNIQUES

DNA as a metrology standard for length and force measurements with optical tweezers

John Peter Rickgauer 1, Derek N Fuller 1 and Douglas Smith 2*

1 University of California, San Diego
2 University of California at San Diego

* To whom correspondence should be addressed. E-mail: des{at}physics.ucsd.edu.

Submitted on May 18, 2006
Revised on June 26, 2006
Accepted on 24 August 2006


  Abstract
Optical tweezers have broad applications in studies of structures and processes in molecular and cellular biophysics. Use of optical tweezers for quantitative molecular-scale measurement requires careful calibration in physical units. Here we show that DNA molecules may be used as metrology standards for force and length measurements. Analysis of DNA molecules of two specific lengths allows simultaneous determination of all essential measurement parameters. We validate this "biological calibration" method experimentally and with simulated data, and show that precisions in determining length scale factor (~0.2%), length offset (~0.03%), force scale factor (~2%), and compliance of the traps (~3%) are limited only by current measurement variation, much of which arises from polydispersity of the microspheres (~2%). We find this procedure to be simpler and more convenient than previous methods, and suggest that it provides an easily replicated standard that can insure uniformity of measurements made in different laboratories.

Key Words: Calibration, DNA, Force, Laser, Optical tweezers, Single-molecule




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Copyright © 2006 by the Biophysical Society.