GSM9474752: THP-1 cells, PAX-8 target genes (ChIP); Homo sapiens; ChIP-Seq
Following C18-3OH treatment, the cells were washed to remove the culture medium and stored at −80°C for subsequent analysis. Frozen cell pellets were thawed and subjected to chromatin immunoprecipitation sequencing (ChIP-Seq) to identify PAX-8 target genes. Cell samples were processed through lysis, sonication, immunoprecipitation, and cross-linking to obtain DNA. The purified DNA fragments, after precipitation and dilution, were used to construct a ChIP-seq library for sequencing. The ChIP-seq experiment was independently repeated once . The experimental procedures were performed by Shanghai Kangcheng Biotechnology Co., Ltd., with standardized protocols executed by trained personnel. ChIP DNA QC: The purity and concentration of DNA samples were determined with Qubit® Fluorometer. Results are provided in this report. Sequencing Library Preparation: DNA samples were end-repaired, A tailed, and adaptor ligated using TruSeq Nano DNASample Prep Kit (#FC-121-4002, Illumina), following the manufacturer's instructions. ~200-1500 bp fragments were size selected using AMPure XP beads. The final size of the library was confirmed by Agilent 2100 Bioanalyzer. Cluster Generation: The samples were diluted to a final concentration of 8 pM and cluster generation was performed on the Illumina cBot following NovaSeq 6000 S4 Reagent Kit manufacturer's instructions.Sequencing on Illumina NovaSeq 6000: Sequencing was performed on Illumina NovaSeq 6000 using NovaSeq 6000 S4 Reagent Kit (300 cycles), according to the manufacturer's instructions.