GSM9688506: Strain Y152, pSer2-RNAPII, biol rep 2, ChIP; Saccharomyces cerevisiae; ChIP-Seq
Cells were fixed with 1% Formaldehyde (FA) for 15 min at 30oC at 230 rpm and were quenched with 128 mM Glycine. Upon lysis, chromatin was sheared in a Covaris Sonicator. pSer2-RNAPII-Immunoprecipitated chromatin was treated with Rnase A and Proteinase K, and DNA was isolated and precipitated with Phenol-Chloroform extraction and Ethanol 100%. ChIP-seq were prepared using NEBNext® Ultra™ II DNA Library Prep with Sample Purification Beads according to manufacturer's protocol. To account for the smaller amounts of immunoprecipitated materials in ChIP, adaptor was diluted 10-fold before adaptor ligation. Libraries were amplified with 3 and 7 cycles of PCR for input and ChIP samples respectively. Fragment size and quantification of amplified libraries were analyzed on fragment analyzer (Agilent) and size selected with QIAseq® Beads if needed before library pooling and sequencing. Libraries were sequenced for 150 cycles in paired-end mode on the Illumina NovaSeq X+ 1.5B platform