title: GSM6659843: GSK458_antiH3K27Ac; Homo sapiens; ChIP-Seq
library_construction_protocol: The cell lysate was sonicated. Protein concentrations of whole-cell lysates were determined using Lowry assays (Bio-Rad). Samples containing 750 µg of protein were used per immunoprecipitation (IP) reaction. Protein of interest was immunoprecipitated using Thermo Fisher Dynabead A kit (cat# 10006D), following the manufacturer's instructions. For immunoprecipitation the H3K27ac antibody (Abcam cat# ab4729) and IgG included in the Diagenode kit was used. Following immunoprecipitation, half of the precipitate Libraries were prepared with the Swift Accel-NGS 2S Plus DNA kit and 2S Combinatorial Dual Indexing kit, according to the manufacturer's instructions. Each library was prepared with 3.9ng of DNA. The final library amplification was performed with 8 cycles of PCR amplification. The Qubit dsDNA HS Assay Kit and High Sensitivity D1000 ScreenTape assay were used for QC assessment. The KAPA Library Quanitification Kit for Illumina platforms (Roche) was used to obtain library concentrations and guide pooling. Libraries were sequenced on the NovaSeq 6000 (Illumina) using the S1 flow cell and paired-end 50-nucleotide chemistry.