| Simultaneous, coincident optical trapping and single-molecule fluorescence. | |
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MedLine Citation:
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PMID: 15782176 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We constructed a microscope-based instrument capable of simultaneous, spatially coincident optical trapping and single-molecule fluorescence. The capabilities of this apparatus were demonstrated by studying the force-induced strand separation of a dye-labeled, 15-base-pair region of double-stranded DNA (dsDNA), with force applied either parallel ('unzipping' mode) or perpendicular ('shearing' mode) to the long axis of the region. Mechanical transitions corresponding to DNA hybrid rupture occurred simultaneously with discontinuous changes in the fluorescence emission. The rupture force was strongly dependent on the direction of applied force, indicating the existence of distinct unbinding pathways for the two force-loading modes. From the rupture force histograms, we determined the distance to the thermodynamic transition state and the thermal off rates in the absence of load for both processes. |
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Authors:
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Matthew J Lang; Polly M Fordyce; Anita M Engh; Keir C Neuman; Steven M Block |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Validation Studies Date: 2004-10-21 |
Journal Detail:
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Title: Nature methods Volume: 1 ISSN: 1548-7091 ISO Abbreviation: Nat. Methods Publication Date: 2004 Nov |
Date Detail:
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Created Date: 2005-05-03 Completed Date: 2005-05-24 Revised Date: 2013-06-09 |
Medline Journal Info:
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Nlm Unique ID: 101215604 Medline TA: Nat Methods Country: United States |
Other Details:
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Languages: eng Pagination: 133-9 Citation Subset: IM |
Affiliation:
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Department of Biological Sciences, Stanford University, Stanford, California 94305, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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DNA
/
analysis*,
chemistry* Elasticity Equipment Design Equipment Failure Analysis Lasers* Micromanipulation / instrumentation*, methods Microscopy, Fluorescence / instrumentation*, methods Microscopy, Video / instrumentation*, methods Physical Stimulation / instrumentation*, methods Stress, Mechanical Systems Integration |
| Grant Support | |
ID/Acronym/Agency:
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R01 GM051453-12/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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9007-49-2/DNA |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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