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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...

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Autores principales: Segorbe, David, Wilkinson, Derek, Mizeranschi, Alexandru, Hughes, Timothy, Aaløkken, Ragnhild, Váchová, Libuše, Palková, Zdena, Gilfillan, Gregor D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
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.
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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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