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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
F1000 Research Limited
2022
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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. |
format | Online Article Text |
id | pubmed-10372461 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | F1000 Research Limited |
record_format | MEDLINE/PubMed |
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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