Chromatin Affinity Purification (ChAP) from Arabidopsis thaliana Rosette Leaves Using in vivo Biotinylation System

Weronika Sura; Piotr A Ziolkowski

doi:10.21769/BioProtoc.2677

Chromatin Affinity Purification (ChAP) from Arabidopsis thaliana Rosette Leaves Using in vivo Biotinylation System

Bio Protoc. 2018 Jan 5;8(1):e2677. doi: 10.21769/BioProtoc.2677.

Authors

Weronika Sura¹, Piotr A Ziolkowski¹

Affiliation

¹ Department of Genome Biology, Adam Mickiewicz University in Poznan, Poznan, Poland.

Abstract

Chromatin Affinity Purification (ChAP) is widely used to study chromatin architecture and protein complexes interacting with DNA. Here we present an efficient method for ChAP from Arabidopsis thaliana rosette leaves, in which in vivo biotinylation system is used. The chromatin is digested by Micrococcal Nuclease (MNase), hence the distribution of nucleosomes is also achieved. The in vivo biotinylation system was initially developed for Drosophila melanogaster ( Mito et al., 2005 ), but the presented protocol has been developed specifically for Arabidopsis thaliana ( Sura et al., 2017 ).

Keywords: Arabidopsis thaliana; ChAP-qPCR; ChIP; Chromatin MNase digestion; Chromatin affinity purification; Chromatin immunoprecipitation; Histone distribution; Nucleosome occupancy.