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.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Anesthesiology and Intensive Care Medicine, Justus-Liebig-University Giessen, Germany; German Centre for Infection Research (DZIF), partner site Giessen-Marburg-Langen, Giessen, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Weiterer
authors.affiliation: Department of Anesthesiology and Intensive Care Medicine, Justus-Liebig-University Giessen, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Uhle
authors.affiliation: Genome Analytics, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Sabin
authors.initials: S
authors.lastname: Bhuju
authors.affiliation: Genome Analytics, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Jarek
authors.affiliation: Department of Anesthesiology and Intensive Care Medicine, Justus-Liebig-University Giessen, Germany; German Centre for Infection Research (DZIF), partner site Giessen-Marburg-Langen, Giessen, Germany.
authors.forename: Markus A
authors.initials: MA
authors.lastname: Weigand
authors.affiliation: Institute for Genetics, Justus-Liebig-University, Giessen, Germany.
authors.forename: Marek
authors.initials: M
authors.lastname: Bartkuhn
mesh_terms: D020029:Base Pairing / Q000235:genetics; D001665:Binding Sites; D002469:Cell Separation; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D002881:Chromosomes, Human, Pair 12 / Q000235:genetics; D019295:Computational Biology / Q000379:methods*; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D009000:Monocytes / Q000166:cytology* / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques; D013223:Statistics as Topic
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