title: GSM2187257: H3K4me3-shJARID1D_ChIP-Seq_rep1; Homo sapiens; ChIP-Seq
library_construction_protocol: ChIP with reference exogenous genome (ChIP-Rx) was performed to compare H3K4me1 and H3K4me3 levels in shLuciferase, shJARID1D or shZMYND8 DU145 human prostate cancer cells. Drosophila melanogaster chromatin and Drosophila-specific H2Av antibody were spiked-in to each ChIP reaction as a minor fraction. Cells were cross-linked with 1% formaldehyde at room temperature for 10 min and then lysed and sonicated to fragment genomic DNA sizes to 200-500 bp. Chromatin samples were incubated with specific antibodies overnight at 4℃. The protein-DNA complexes were immobilized on protein A/G beads. The bound fractions were washed with low salt, high salt, LiCI wash buffer and TE buffer. Reverse-crosslinking of the eluted DNA were carried out at 67℃ for 8 hours. After RNase A and protease K digestion, DNA fraction was purified using PCR purification kit. ChIP DNA libraries were prepared for sequencing using standard Illumina protocols.