GSM8578848: pht1-K84H-input-2; Schizosaccharomyces pombe; ChIP-Seq
Cells were fixed with 1% formaldehyde for 10 minutes. After quenching and cell lysis with beads via a Precellys homogenizer, chromatin was sonicated with a E220 focused ultra-sonicator (Covaris). ChIP-seq libraries were prepared using the NEBNext Ultra II library preparation kit for Illumina complemented with custom synthesized barcoded sequencing adapters provided by the Vienna BioCenter Core Facilities Next Generation Sequencing Facility. PCR was performed as a 50 µl reaction using the Q5 polymerase (NEB). Samples were PCR-amplified to the beginning of the logarithmic phase, as determined using a small-scale qPCR reaction run in parallel (typically around 8-10 cycles). Following quality control by Agilent Fragment Analyzer and quantification by RT-qPCR using a Kapa Library Quantification Kit, libraries were sequenced at the Vienna BioCenter Core Facilities Next Generation Sequencing Core Facility on an Illumina NovaSeq X.