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Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni

In living cells, the genetic information stored in the DNA sequence is always associated with chromosomal and extra-chromosomal epigenetic information. Chromatin is formed by the DNA and associated proteins, in particular histones. Covalent histone modifications are important bearers of epigenetic i...

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Autores principales: Lasica, Chrystelle, de Carvalho Augusto, Ronaldo, Moné, Hélène, Mouahid, Gabriel, Chaparro, Cristian, Veillard, Anne-Clémence, Zelisko-Schmidt, Agnieszka, Grunau, Christoph
Formato: Online Artículo Texto
Lenguaje:English
Publicado: F1000 Research Limited 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10372461/
https://www.ncbi.nlm.nih.gov/pubmed/37521535
http://dx.doi.org/10.12688/wellcomeopenres.17779.1
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author Lasica, Chrystelle
de Carvalho Augusto, Ronaldo
Moné, Hélène
Mouahid, Gabriel
Chaparro, Cristian
Veillard, Anne-Clémence
Zelisko-Schmidt, Agnieszka
Grunau, Christoph
author_facet Lasica, Chrystelle
de Carvalho Augusto, Ronaldo
Moné, Hélène
Mouahid, Gabriel
Chaparro, Cristian
Veillard, Anne-Clémence
Zelisko-Schmidt, Agnieszka
Grunau, Christoph
author_sort Lasica, Chrystelle
collection PubMed
description In living cells, the genetic information stored in the DNA sequence is always associated with chromosomal and extra-chromosomal epigenetic information. Chromatin is formed by the DNA and associated proteins, in particular histones. Covalent histone modifications are important bearers of epigenetic information and as such have been increasingly studied since about the year 2000. One of the principal techniques to gather information about the association between DNA and modified histones is chromatin immunoprecipitation (ChIP), also combined with massive sequencing (ChIP-Seq). Automated ChIPmentation procedure is a convenient alternative to native chromatin immunoprecipitation (N-ChIP). It is now routinely used for ChIP-Seq in many model species, using in general roughly 10 (6) cells per experiment. Such high cell numbers are sometimes difficult to produce. Using the human parasite Schistosoma mansoni, whose production requires sacrificing animals and should therefore be kept to a minimum, we show here that automated ChIPmentation is suitable for limited biological material. We define the operational limit as ≥20,000 Schistosoma cells. We also present a streamlined protocol for the preparation of ChIP input libraries.
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spelling pubmed-103724612023-07-28 Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni Lasica, Chrystelle de Carvalho Augusto, Ronaldo Moné, Hélène Mouahid, Gabriel Chaparro, Cristian Veillard, Anne-Clémence Zelisko-Schmidt, Agnieszka Grunau, Christoph Wellcome Open Res Method Article In living cells, the genetic information stored in the DNA sequence is always associated with chromosomal and extra-chromosomal epigenetic information. Chromatin is formed by the DNA and associated proteins, in particular histones. Covalent histone modifications are important bearers of epigenetic information and as such have been increasingly studied since about the year 2000. One of the principal techniques to gather information about the association between DNA and modified histones is chromatin immunoprecipitation (ChIP), also combined with massive sequencing (ChIP-Seq). Automated ChIPmentation procedure is a convenient alternative to native chromatin immunoprecipitation (N-ChIP). It is now routinely used for ChIP-Seq in many model species, using in general roughly 10 (6) cells per experiment. Such high cell numbers are sometimes difficult to produce. Using the human parasite Schistosoma mansoni, whose production requires sacrificing animals and should therefore be kept to a minimum, we show here that automated ChIPmentation is suitable for limited biological material. We define the operational limit as ≥20,000 Schistosoma cells. We also present a streamlined protocol for the preparation of ChIP input libraries. F1000 Research Limited 2022-04-11 /pmc/articles/PMC10372461/ /pubmed/37521535 http://dx.doi.org/10.12688/wellcomeopenres.17779.1 Text en Copyright: © 2022 Lasica C et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Method Article
Lasica, Chrystelle
de Carvalho Augusto, Ronaldo
Moné, Hélène
Mouahid, Gabriel
Chaparro, Cristian
Veillard, Anne-Clémence
Zelisko-Schmidt, Agnieszka
Grunau, Christoph
Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni
title Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni
title_full Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni
title_fullStr Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni
title_full_unstemmed Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni
title_short Automated ChIPmentation procedure on limited biological material of the human blood fluke Schistosoma mansoni
title_sort automated chipmentation procedure on limited biological material of the human blood fluke schistosoma mansoni
topic Method Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10372461/
https://www.ncbi.nlm.nih.gov/pubmed/37521535
http://dx.doi.org/10.12688/wellcomeopenres.17779.1
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