title: GSM9058355: Ser5p Pol II ChIP-seq, 4h auxin, replicate 1; Homo sapiens; ChIP-Seq
library_name: GSM9058355
library_construction_protocol: The cells were lysed to prepare nuclear extracts. Following chromatin shearing by sonication, the lysates were incubated overnight at 4°C with protein Dynabeads™ M-280 Sheep Anti-Mouse IgG or Anti-Rabbit IgG (Invitrogen 11202D, 11203D) coupled with 10μg of the following antibodies: anti-FLAG M2 (Sigma F3165), anti-Pol II Ser5p (Abcam Ab5131) and anti-Pol II Ser2p (Diagenode C15200005) for endogenous ZC3H4 and Pol II purification, respectively. Following immunoprecipitation, the beads were retrieved using a magnet, washed, and the chromatin was eluted and de-crosslinked overnight at 65°C. DNA was subsequently purified using QIAquick PCR Purification Kit (Qiagen  28104) following the manufacturer instructions, and its quantity was determined using QuantiFluor (Promega) Library preparation for Illumina sequencing was carried out using a described protocol (PMID: 29706544). The quality and quantity of the purified DNA libraries were assessed using Tapestation (Agilent Technologies) and Qubit (Life Technologies) before being subjected to sequencing on the NovaSeq platform