GSM4879831: ChIP-seq input DNA: Spt6-FLAG background: spt6-YW pob3-E154K, 37C, 1; Saccharomyces cerevisiae; ChIP-Seq
The cultures were processed for cross-linking and collecting as previously described (Doris et al., 2018). Chromatin was prepared using standard methods (Gopalakrishnan et al., 2019). Each chromatin sample was mixed with S. pombe chromatin (strain FWP570) at 10% level by protein mass for spike-in normalization and split into aliquots of 500 µg of chromatin for immunoprecipitation. Chromatin precipitations were performed overnight at 4°C using either 50 µl of anti-FLAG M2 affinity gel (Sigma) for Spt6, 30 µl of anti-Myc antibodies (9E10, Santa Cruz Biotechnology), 8 µl of 8WG16 antibodies (Millipore Sigma) for Rpb1, or 5 µl of anti-V5 antibodies (Invitrogen) per 500 µg of chromatin. DNA isolated from each sample before precipitation (inputs) and after precipitation (IP) was used for library generation as previously described (Gopalakrishnan et al., 2019).