AI Article Synopsis
- The study examines single-molecule DNA digestion using exonuclease III, which works in a 3' to 5' direction, in a micro-channel with two layers of laminar flow.
- A DNA-bead complex was trapped and stretched in the absence of exonuclease III, then moved to a flow layer containing the enzyme, initiating the digestion process.
- The reaction revealed that the DNA shortened linearly over time, and the single-molecule digestion rate was about 28 times faster than what was observed in a bulk experiment.
Article Abstract
Single-molecule DNA digestion by exonuclease III, which has 3' to 5' exonuclease activity, was analyzed using a micro-channel with two-layer laminar flow. First, a DNA-bead complex was optically trapped in one layer in the absence of exonuclease III permitted the DNA to be stretched by the laminar flow. The exonuclease III reaction was initiated by moving the trapped DNA-bead complex to another layer of flow, which contained exonuclease III. As the reaction proceeded, the fluorescently-stained DNA was observed to shorten. The process was photographed; examination of the photographs showed that the DNA molecule shortened in a linear fashion with respect to the reaction time. The digestion rate obtained from the single-molecule experiment was compared to that measured from a bulk experiment and was found to be ca. 28 times higher than the bulk digestion rate.
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| http://dx.doi.org/10.1080/07391102.2008.10507194 | DOI Listing |
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