GSM1576448: AR LNCaP Input; Homo sapiens; ChIP-Seq
LNCaP cells were starved in phenol-free RPMI medium supplemented with 10% charcoal-stripped FBS for 3 days followed by treatment with 10nM DHT or 100nM calcitriol for 1hr. Cells were fixed in 1% formaldehyde for 15min and quenched with 0.125M glycine. Chromatin was isolated with lysis buffer and glass homogenization, then sonicated by Bioruptor (Diagenode). For each ChIP, An aliquot of 50 ug chromatin was pre-cleared with BSA-blocked magnetic protein G beads at 4°C  overnight (Millipore). 5ug antibody was added to pre-cleared chromatin and incubated at 4°C for 6 hrs. BSA-blocked beads were then added and incubated at 4°C overnight. Samples were then washed 4 times with washing buffer and 2 times with TE, followed by Rnase A treatment for 2hr and proteinase K treatment for 2hr. Crosslinks were reversed by incubation overnight at 65 C. Finally, ChIPped DNA was extracted by MinElute PCR Purification kit (Qiagen). DNA samples after ChIP were made into libraries with TrueSeq DNA Sample Prep Kit (Illumina) and run on Illumina Hiseq 2000 to generate single end 50bp reads.
GEO
GSM1576448