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dc.contributor.authorWeiterer, Sebastian
dc.contributor.authorUhle, Florian
dc.contributor.authorBhuju, Sabin
dc.contributor.authorJarek, Michael
dc.contributor.authorWeigand, Markus A
dc.contributor.authorBartkuhn, Marek
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.
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
refterms.dateFOA2018-06-13T01:36:47Z
html.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.


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