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A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules

We report the development of a magnetic tweezers that can be used to micromanipulate single DNA molecules by applying picoNewton (pN)-scale forces in the horizontal plane. The resulting force–extension data from our experiments show high-resolution detection of changes in the DNA tether’s extension:...

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Detalles Bibliográficos
Autores principales: Fabian, Roberto, Tyson, Christopher, Tuma, Pamela L., Pegg, Ian, Sarkar, Abhijit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187538/
https://www.ncbi.nlm.nih.gov/pubmed/30424121
http://dx.doi.org/10.3390/mi9040188
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author Fabian, Roberto
Tyson, Christopher
Tuma, Pamela L.
Pegg, Ian
Sarkar, Abhijit
author_facet Fabian, Roberto
Tyson, Christopher
Tuma, Pamela L.
Pegg, Ian
Sarkar, Abhijit
author_sort Fabian, Roberto
collection PubMed
description We report the development of a magnetic tweezers that can be used to micromanipulate single DNA molecules by applying picoNewton (pN)-scale forces in the horizontal plane. The resulting force–extension data from our experiments show high-resolution detection of changes in the DNA tether’s extension: ~0.5 pN in the force and <10 nm change in extension. We calibrate our instrument using multiple orthogonal techniques including the well-characterized DNA overstretching transition. We also quantify the repeatability of force and extension measurements, and present data on the behavior of the overstretching transition under varying salt conditions. The design and experimental protocols are described in detail, which should enable straightforward reproduction of the tweezers.
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spelling pubmed-61875382018-11-01 A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules Fabian, Roberto Tyson, Christopher Tuma, Pamela L. Pegg, Ian Sarkar, Abhijit Micromachines (Basel) Article We report the development of a magnetic tweezers that can be used to micromanipulate single DNA molecules by applying picoNewton (pN)-scale forces in the horizontal plane. The resulting force–extension data from our experiments show high-resolution detection of changes in the DNA tether’s extension: ~0.5 pN in the force and <10 nm change in extension. We calibrate our instrument using multiple orthogonal techniques including the well-characterized DNA overstretching transition. We also quantify the repeatability of force and extension measurements, and present data on the behavior of the overstretching transition under varying salt conditions. The design and experimental protocols are described in detail, which should enable straightforward reproduction of the tweezers. MDPI 2018-04-17 /pmc/articles/PMC6187538/ /pubmed/30424121 http://dx.doi.org/10.3390/mi9040188 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fabian, Roberto
Tyson, Christopher
Tuma, Pamela L.
Pegg, Ian
Sarkar, Abhijit
A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules
title A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules
title_full A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules
title_fullStr A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules
title_full_unstemmed A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules
title_short A Horizontal Magnetic Tweezers and Its Use for Studying Single DNA Molecules
title_sort horizontal magnetic tweezers and its use for studying single dna molecules
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187538/
https://www.ncbi.nlm.nih.gov/pubmed/30424121
http://dx.doi.org/10.3390/mi9040188
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