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Single molecule mass photometry of nucleic acids
Mass photometry is a recently developed methodology capable of measuring the mass of individual proteins under solution conditions. Here, we show that this approach is equally applicable to nucleic acids, enabling their facile, rapid and accurate detection and quantification using sub-picomoles of s...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7515692/ https://www.ncbi.nlm.nih.gov/pubmed/32756898 http://dx.doi.org/10.1093/nar/gkaa632 |
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author | Li, Yiwen Struwe, Weston B Kukura, Philipp |
author_facet | Li, Yiwen Struwe, Weston B Kukura, Philipp |
author_sort | Li, Yiwen |
collection | PubMed |
description | Mass photometry is a recently developed methodology capable of measuring the mass of individual proteins under solution conditions. Here, we show that this approach is equally applicable to nucleic acids, enabling their facile, rapid and accurate detection and quantification using sub-picomoles of sample. The ability to count individual molecules directly measures relative concentrations in complex mixtures without need for separation. Using a dsDNA ladder, we find a linear relationship between the number of bases per molecule and the associated imaging contrast for up to 1200 bp, enabling us to quantify dsDNA length with up to 2 bp accuracy. These results introduce mass photometry as an accurate, rapid and label-free single molecule method complementary to existing DNA characterization techniques. |
format | Online Article Text |
id | pubmed-7515692 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-75156922020-09-30 Single molecule mass photometry of nucleic acids Li, Yiwen Struwe, Weston B Kukura, Philipp Nucleic Acids Res Methods Online Mass photometry is a recently developed methodology capable of measuring the mass of individual proteins under solution conditions. Here, we show that this approach is equally applicable to nucleic acids, enabling their facile, rapid and accurate detection and quantification using sub-picomoles of sample. The ability to count individual molecules directly measures relative concentrations in complex mixtures without need for separation. Using a dsDNA ladder, we find a linear relationship between the number of bases per molecule and the associated imaging contrast for up to 1200 bp, enabling us to quantify dsDNA length with up to 2 bp accuracy. These results introduce mass photometry as an accurate, rapid and label-free single molecule method complementary to existing DNA characterization techniques. Oxford University Press 2020-08-05 /pmc/articles/PMC7515692/ /pubmed/32756898 http://dx.doi.org/10.1093/nar/gkaa632 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methods Online Li, Yiwen Struwe, Weston B Kukura, Philipp Single molecule mass photometry of nucleic acids |
title | Single molecule mass photometry of nucleic acids |
title_full | Single molecule mass photometry of nucleic acids |
title_fullStr | Single molecule mass photometry of nucleic acids |
title_full_unstemmed | Single molecule mass photometry of nucleic acids |
title_short | Single molecule mass photometry of nucleic acids |
title_sort | single molecule mass photometry of nucleic acids |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7515692/ https://www.ncbi.nlm.nih.gov/pubmed/32756898 http://dx.doi.org/10.1093/nar/gkaa632 |
work_keys_str_mv | AT liyiwen singlemoleculemassphotometryofnucleicacids AT struwewestonb singlemoleculemassphotometryofnucleicacids AT kukuraphilipp singlemoleculemassphotometryofnucleicacids |