Journalartikel

Jahr der Veröffentlichung: 2014

Zeitschrift: PLoS ONE

Bandnummer: 9

Heftnummer: 4

ISSN: 1932-6203

Open Access Status: Gold

DOI Link: https://doi.org/10.1371/journal.pone.0094164

Verlag: Public Library of Science

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.

Harvard-Zitierstil: Weiterer, S., Uhle, F., Bhuju, S., Jarek, M., Weigand, M. and Bartkuhn, M. (2014) 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, PLoS ONE, 9(4), Article e94164. https://doi.org/10.1371/journal.pone.0094164

APA-Zitierstil: Weiterer, S., Uhle, F., Bhuju, S., Jarek, M., Weigand, M., & Bartkuhn, M. (2014). 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. PLoS ONE. 9(4), Article e94164. https://doi.org/10.1371/journal.pone.0094164