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An optimized FAIRE procedure for low cell numbers in yeast
We report an optimized low‐input FAIRE‐seq (Formaldehyde‐Assisted Isolation of Regulatory Elements‐sequencing) procedure to assay chromatin accessibility from limited amounts of yeast cells. We demonstrate that the method performs well on as little as 4 mg of cells scraped directly from a few coloni...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6099244/ https://www.ncbi.nlm.nih.gov/pubmed/29577419 http://dx.doi.org/10.1002/yea.3316 |
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author | Segorbe, David Wilkinson, Derek Mizeranschi, Alexandru Hughes, Timothy Aaløkken, Ragnhild Váchová, Libuše Palková, Zdena Gilfillan, Gregor D. |
author_facet | Segorbe, David Wilkinson, Derek Mizeranschi, Alexandru Hughes, Timothy Aaløkken, Ragnhild Váchová, Libuše Palková, Zdena Gilfillan, Gregor D. |
author_sort | Segorbe, David |
collection | PubMed |
description | We report an optimized low‐input FAIRE‐seq (Formaldehyde‐Assisted Isolation of Regulatory Elements‐sequencing) procedure to assay chromatin accessibility from limited amounts of yeast cells. We demonstrate that the method performs well on as little as 4 mg of cells scraped directly from a few colonies. Sensitivity, specificity and reproducibility of the scaled‐down method are comparable with those of regular, higher input amounts, and allow the use of 100‐fold fewer cells than existing procedures. The method enables epigenetic analysis of chromatin structure without the need for cell multiplication of exponentially growing cells in liquid culture, thus opening the possibility of studying colony cell subpopulations, or those that can be isolated directly from environmental samples. |
format | Online Article Text |
id | pubmed-6099244 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-60992442018-08-23 An optimized FAIRE procedure for low cell numbers in yeast Segorbe, David Wilkinson, Derek Mizeranschi, Alexandru Hughes, Timothy Aaløkken, Ragnhild Váchová, Libuše Palková, Zdena Gilfillan, Gregor D. Yeast Genome Screen Reports We report an optimized low‐input FAIRE‐seq (Formaldehyde‐Assisted Isolation of Regulatory Elements‐sequencing) procedure to assay chromatin accessibility from limited amounts of yeast cells. We demonstrate that the method performs well on as little as 4 mg of cells scraped directly from a few colonies. Sensitivity, specificity and reproducibility of the scaled‐down method are comparable with those of regular, higher input amounts, and allow the use of 100‐fold fewer cells than existing procedures. The method enables epigenetic analysis of chromatin structure without the need for cell multiplication of exponentially growing cells in liquid culture, thus opening the possibility of studying colony cell subpopulations, or those that can be isolated directly from environmental samples. John Wiley and Sons Inc. 2018-05-28 2018-08 /pmc/articles/PMC6099244/ /pubmed/29577419 http://dx.doi.org/10.1002/yea.3316 Text en © 2018 The Authors. Yeast published by John Wiley & Sons, Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Genome Screen Reports Segorbe, David Wilkinson, Derek Mizeranschi, Alexandru Hughes, Timothy Aaløkken, Ragnhild Váchová, Libuše Palková, Zdena Gilfillan, Gregor D. An optimized FAIRE procedure for low cell numbers in yeast |
title | An optimized FAIRE procedure for low cell numbers in yeast |
title_full | An optimized FAIRE procedure for low cell numbers in yeast |
title_fullStr | An optimized FAIRE procedure for low cell numbers in yeast |
title_full_unstemmed | An optimized FAIRE procedure for low cell numbers in yeast |
title_short | An optimized FAIRE procedure for low cell numbers in yeast |
title_sort | optimized faire procedure for low cell numbers in yeast |
topic | Genome Screen Reports |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6099244/ https://www.ncbi.nlm.nih.gov/pubmed/29577419 http://dx.doi.org/10.1002/yea.3316 |
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