GSM4367475
GSM4367475: Mg-Ecs-IP; Homo sapiens; ChIP-Seq
Briefly, HCAEC were collected from seeding on Mg-based alloy discs after 2 days. Cells were then fixed with1% formaldehyde (Sigma) in medium for 10min at RT, and lysed in cell lysis buffer. Chromatin was sheared to approximately 200bp fragments by using ULTRASONIC PROCESSOR (Fisher Scientific, USA). For ChIP-seq, sheared chromatin was incubated with anti-YAP antibody overnight at 4°C. Antibody/antigen complexes were recovered with ProteinA-Dynabeads (Invitrogen) for 2 h at 4 °C. 10ng ChIP'd DNA of each sample was converted to phosphorylated blunt-ended; An 'A' base was added to the 3' end of the blunt phosphorylated DNA fragments; Illumina's genomic adapters were ligated to the A tailed DNA fragments; PCR amplification was performed to enrich ligated fragments. The enriched product of ~200-1500 bp was size selected using AMPure XP beads. The library was denatured with 0.1M NaOH to generate single-stranded DNA molecules, and loaded onto channels of the flow cell at 8pM concentration, amplified in situ using HiSeq 3000/4000 PE Cluster Kit (Illumina). Sequencing was carried out by running 2× 150 cycles on Illumina HiSeq 4000 according to the manufacturer's instructions. DNA samples were end-repaired, A tailed, and adaptor ligated using TruSeq Nano DNA Sample 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.
GEO
GSM4367475
GEO Accession
GSM4367475