GSM5195594: IgG ChIP-seq rep1; Homo sapiens; ChIP-Seq
For biotin ChIP-seq, cells were crosslinked with 1% PFA, permeabilized and incubated with an a-H3K9me3 or a-IgG antibody, followed by binding of protA-Turbo, biotinylation for 10 minutes, lysis and IP with magnetic streptavidin beads. For flywch1 and IgG ChIP-seq, cells were crosslinked in 1% PFA and chromatin was sonicated to 300-800 bp fragments and IPs were performed with antibodies against V5 (present in the flywch1-V5-GFP fusion protein) or IgG. Libraries were prepared using the KAPA HyperPrep Kit (Kapa Biosystems) according to the manufacturer's instructions, which some minor modifications. There include 2 ng DNA as input material for the library preparation, 12 PCR cycles for the library amplification, and a 0.65x reverse phase size selection followed by a double 1.0x bead-based clean up after library amplification. Library concentrations were determined using the DeNovix dsDNA High Sensitivity Assay (DeNovix), and fragment distribution using a High Sensitivity DNA Chip on a Bioanalyzer 2100 system (Aligent Technologies).