Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexes
We report the development of a simple-to-implement magnetic force transducer that can apply a wide range of piconewton (pN) scale forces on single DNA molecules and DNA—protein complexes in the horizontal plane. The resulting low-noise force-extension data enable very high-resolution detection of ch...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2016-01-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/000114369 |
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| author | Christopher P. McAndrew Christopher Tyson Joseph Zischkau Patrick Mehl Pamela L. Tuma Ian L. Pegg Abhijit Sarkar |
| author_facet | Christopher P. McAndrew Christopher Tyson Joseph Zischkau Patrick Mehl Pamela L. Tuma Ian L. Pegg Abhijit Sarkar |
| author_sort | Christopher P. McAndrew |
| collection | DOAJ |
| description | We report the development of a simple-to-implement magnetic force transducer that can apply a wide range of piconewton (pN) scale forces on single DNA molecules and DNA—protein complexes in the horizontal plane. The resulting low-noise force-extension data enable very high-resolution detection of changes in the DNA tether's extension: ∼0.05 pN in force and <10 nm change in extension. We have also verified that we can manipulate DNA in near equilibrium conditions through the wide range of forces by ramping the force from low to high and back again, and observing minimal hysteresis in the molecule's force response. Using a calibration technique based on Stokes' drag law, we have confirmed our force measurements from DNA force-extension experiments obtained using the fluctuation-dissipation theorem applied to transverse fluctuations of the magnetic microsphere. We present data on the force-distance characteristics of a DNA molecule complexed with histones. The results illustrate how the tweezers can be used to study DNA binding proteins at the single molecule level. |
| format | Article |
| id | doaj-art-3ca6bd1ff55245769c8b33664ded7561 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-3ca6bd1ff55245769c8b33664ded75612025-08-20T02:25:58ZengTaylor & Francis GroupBioTechniques0736-62051940-98182016-01-01601212710.2144/000114369Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexesChristopher P. McAndrew0Christopher Tyson1Joseph Zischkau2Patrick Mehl3Pamela L. Tuma4Ian L. Pegg5Abhijit Sarkar61Department of Physics, The Catholic University of America, Washington, DC2Vitreous State Laboratory, Washington, DC3Carnegie Mellon University, Pittsburgh, PA2Vitreous State Laboratory, Washington, DC4Department of Biology, The Catholic University of America, Washington, DC1Department of Physics, The Catholic University of America, Washington, DC1Department of Physics, The Catholic University of America, Washington, DCWe report the development of a simple-to-implement magnetic force transducer that can apply a wide range of piconewton (pN) scale forces on single DNA molecules and DNA—protein complexes in the horizontal plane. The resulting low-noise force-extension data enable very high-resolution detection of changes in the DNA tether's extension: ∼0.05 pN in force and <10 nm change in extension. We have also verified that we can manipulate DNA in near equilibrium conditions through the wide range of forces by ramping the force from low to high and back again, and observing minimal hysteresis in the molecule's force response. Using a calibration technique based on Stokes' drag law, we have confirmed our force measurements from DNA force-extension experiments obtained using the fluctuation-dissipation theorem applied to transverse fluctuations of the magnetic microsphere. We present data on the force-distance characteristics of a DNA molecule complexed with histones. The results illustrate how the tweezers can be used to study DNA binding proteins at the single molecule level.https://www.future-science.com/doi/10.2144/000114369single molecule—micromanipulationmagnetic tweezersdifferential measurementmagnetic trap |
| spellingShingle | Christopher P. McAndrew Christopher Tyson Joseph Zischkau Patrick Mehl Pamela L. Tuma Ian L. Pegg Abhijit Sarkar Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexes BioTechniques single molecule—micromanipulation magnetic tweezers differential measurement magnetic trap |
| title | Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexes |
| title_full | Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexes |
| title_fullStr | Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexes |
| title_full_unstemmed | Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexes |
| title_short | Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA—protein complexes |
| title_sort | simple horizontal magnetic tweezers for micromanipulation of single dna molecules and dna protein complexes |
| topic | single molecule—micromanipulation magnetic tweezers differential measurement magnetic trap |
| url | https://www.future-science.com/doi/10.2144/000114369 |
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