From Human Monocytes to Genome-Wide Binding Sites - A Protocol for Small Amounts of Blood: Monocyte Isolation/ChIP-Protocol/Library Amplification/Genome Wide Computational Data Analysis.

2.50
Hdl Handle:
http://hdl.handle.net/10033/317237
Title:
From Human Monocytes to Genome-Wide Binding Sites - A Protocol for Small Amounts of Blood: Monocyte Isolation/ChIP-Protocol/Library Amplification/Genome Wide Computational Data Analysis.
Authors:
Weiterer, Sebastian; Uhle, Florian; Bhuju, Sabin; Jarek, Michael; Weigand, Markus A; Bartkuhn, Marek
Abstract:
Chromatin immunoprecipitation in combination with a genome-wide analysis via high-throughput sequencing is the state of the art method to gain genome-wide representation of histone modification or transcription factor binding profiles. However, chromatin immunoprecipitation analysis in the context of human experimental samples is limited, especially in the case of blood cells. The typically extremely low yields of precipitated DNA are usually not compatible with library amplification for next generation sequencing. We developed a highly reproducible protocol to present a guideline from the first step of isolating monocytes from a blood sample to analyse the distribution of histone modifications in a genome-wide manner. Conclusion: The protocol describes the whole work flow from isolating monocytes from human blood samples followed by a high-sensitivity and small-scale chromatin immunoprecipitation assay with guidance for generating libraries compatible with next generation sequencing from small amounts of immunoprecipitated DNA.
Citation:
From Human Monocytes to Genome-Wide Binding Sites - A Protocol for Small Amounts of Blood: Monocyte Isolation/ChIP-Protocol/Library Amplification/Genome Wide Computational Data Analysis. 2014, 9 (4):e94164 PLoS ONE
Journal:
PloS one
Issue Date:
2014
URI:
http://hdl.handle.net/10033/317237
DOI:
10.1371/journal.pone.0094164
PubMed ID:
24732314
Type:
Article
Language:
en
ISSN:
1932-6203
Appears in Collections:
Publications of Dept. Genome Analysis (GNA)

Full metadata record

DC FieldValue Language
dc.contributor.authorWeiterer, Sebastianen
dc.contributor.authorUhle, Florianen
dc.contributor.authorBhuju, Sabinen
dc.contributor.authorJarek, Michaelen
dc.contributor.authorWeigand, Markus Aen
dc.contributor.authorBartkuhn, Mareken
dc.date.accessioned2014-05-21T13:00:03Z-
dc.date.available2014-05-21T13:00:03Z-
dc.date.issued2014-
dc.identifier.citationFrom Human Monocytes to Genome-Wide Binding Sites - A Protocol for Small Amounts of Blood: Monocyte Isolation/ChIP-Protocol/Library Amplification/Genome Wide Computational Data Analysis. 2014, 9 (4):e94164 PLoS ONEen
dc.identifier.issn1932-6203-
dc.identifier.pmid24732314-
dc.identifier.doi10.1371/journal.pone.0094164-
dc.identifier.urihttp://hdl.handle.net/10033/317237-
dc.description.abstractChromatin immunoprecipitation in combination with a genome-wide analysis via high-throughput sequencing is the state of the art method to gain genome-wide representation of histone modification or transcription factor binding profiles. However, chromatin immunoprecipitation analysis in the context of human experimental samples is limited, especially in the case of blood cells. The typically extremely low yields of precipitated DNA are usually not compatible with library amplification for next generation sequencing. We developed a highly reproducible protocol to present a guideline from the first step of isolating monocytes from a blood sample to analyse the distribution of histone modifications in a genome-wide manner. Conclusion: The protocol describes the whole work flow from isolating monocytes from human blood samples followed by a high-sensitivity and small-scale chromatin immunoprecipitation assay with guidance for generating libraries compatible with next generation sequencing from small amounts of immunoprecipitated DNA.en
dc.language.isoenen
dc.rightsArchived with thanks to PloS oneen
dc.titleFrom Human Monocytes to Genome-Wide Binding Sites - A Protocol for Small Amounts of Blood: Monocyte Isolation/ChIP-Protocol/Library Amplification/Genome Wide Computational Data Analysis.en
dc.typeArticleen
dc.identifier.journalPloS oneen

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