title: GSM4671820: atxr5/atxr6/gcn5 H3K36ac rep2 (ChIPseq); Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: Leaves from three-week-old plants were fixed in 1% formaldehyde. Immunoprecipitation was performed using protein A magnetic beads (New England BioLabs). Following the Proteinase K treatment of each sample, immunoprecipitated DNA was purified using ChIP DNA Clean & Concentrator kit (Zymo Research, Irvine, CA, USA). 2 μl of Histone H3 antibody (Abcam: ab1791), 2.5 μl of H3K27me1 antibody (Millipore: 07-448), 2 ul of H3K27me2 antibody (Abcam: ab24684) or 2.5 μl of H3K27Ac antibody (Active Motif: 39135), was used per immunoprecipitation. For the ChIP experiments for H3K27ac, ChIP with exogenous genome (ChIP-Rx) was performed in order to properly normalize the data (Orlando et al, 2014). For each sample, an equal amount of drosophila chromatin (Active Motif #53083) was added prior to chromatin shearing. ChIP library preparation was performed using TruSeq Library Prep Kit (Illumina). Libraries were validated using Agilent Bioanalyzer 2100 Hisense DNA assay and quantified using the KAPA Library Quantification Kit for Illumina® Platforms kit. Sequencing was done on an Illumina NovaSeq 6000 using the S4 XP workflow.
identifiers.namespace: GEO
identifiers.id: GSM4671820