GSM5738191: Y537S-DoxE2 [ChIP-Seq]; Homo sapiens; ChIP-Seq
The cells on two 100 mm plates were fixed and nuclear pellets were sonicated to prepare soluble chromatin lysate. ER alpha-chromatin complex was immunoprecipitated by anti-ER alpha antibody from the lysate and precipitated DNA was recovered by reverse crosslinking. The precipitated DNA was purified with AMPure XP beads (Agencourt) and ChIP-seq library was prepared from 2 ng of the purified DNA using SMARTer ThruPLEX DNA-seq Kit (TAKARA). The library was then purified using AMPure XP beads (Agencourt) again. The concentration of the library was quantified using Fluorometer (Quantus) and KAPA library quantification kit (Roche), and the quality was checked by TapeStation D5000 (Agilent). The pooled libraries with unique index sequences were sequenced using NextSeq550 (Illumina) with pair-end 75 bp reads following the manufacturer's protocols.