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. 2016 May 24;44(12):5849–5860. doi: 10.1093/nar/gkw445

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Published by Oxford University Press on behalf of Nucleic Acids Research 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

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Figure 1. — RecBCD motion shows variably sized discrete translocations. (A) Precision optical-trapping assay measures RecBCD motion. A DNA molecule is stretched between the RecBCD helicase and an optically trapped bead. One detection laser (pink) measures the bead position (x_bd), which is used to calculate DNA length. The other detection laser (green) measures the position of a fiducial mark (x_fid, y_fid, z_fid), which is used to stabilize the surface in three dimensions. (B) DNA contour length (L) decreases as RecBCD moves forward. Traces taken at 6 pN of force, 2 μM ATP and smoothed to 10 Hz (light green) and 1 Hz (dark green). (C) Magnification of the record shown in (B) (pink box) shows forward motion fit by a step-finding algorithm (55) (black line, arrows) that finds variably sized discrete translocations. (D) Occasionally, sets of discrete backward steps (∼2–3) were observed followed by subsequent forward motion.