Chromatin immunoprecipitation (ChIP) assays were performed in prostate cancer cell line VCaP. To induce the AR activity prior to ChIP, 80% confluent VCaP cells were treated with 1nM of the synthetic androgen methyltrienolone (R1881, dissolved in ethanol) for 24 hours. The ChIPed DNA was blunted and phosphorylated with T4 DNA polymerase, Klenow DNA polymerase and T4 polynucleotide kinase. Subsequently, the DNA fragments was added an A nucleotide to the 3’ end using Klenow exo- (3’ to 5’ exonuclease activity minus). Illumina/Solexa amplification and sequencing adapters were ligated to the DNA-fragments and were size-selected on 2% agarose gel for 150-450bp fragments. After enriched by 16 cycles of PCR amplification, DNA fragments were size-selected again on a 2% agarose gel (150- 450 bp). Purified DNA (Qiagen gel purification kit) was quantified (Nanodrop 1000 spectrophotometer) and subjected to massively parallel sequencing (Illumina platform).