GSM7992962: Brdu-ip H3-HA/T7 WT experiment 1; Saccharomyces cerevisiae; OTHER
We performed eSPAN according to the protocol in our previous studies with some modifications. Briefly, yeast cells were synchronized with alpha-factor and released in YPD medium with BrdU (400ug/ml) following the standard protocol. The cells were digested with MNase (NEB, Cat# M0247S), after saving 1% for BrdU-IP, the rest of digested chromatin was subjected to ChIP against H3K4me3 (ab8580), H3K56ac, HA-Tag (BETHYL,Cat#12CA5), or T7-Tag (BETHYL,Cat#A190-117A), individually. Then DNA extracted from the ChIP products was end-repaired and ligated with adapters using the VAHTS® Universal DNA Library Prep Kit for Illumina V3 (Vazyme, Cat#ND607). After saving 1% for ChIP-seq analysis, the products was subjected to BrdU-IP as previous reported, and the products was amplified with NEBNext High-Fidelity 2X PCR Master Mix (NEB, Cat# M0541L) and indexing primers. The obtained libraries were sequenced using the paired-end method by Illumina nova 6000. eSPAN