GSM3179301: Capture-C for IGHM enhancer; Homo sapiens; OTHER
Three oligos were designed near the cutting sites to capture IGH enhancer (Eμ). Nalm6 was cultured to about 80% confluence and resuspended 10^7 cells in 10 ml of medium. Cells were crosslinked with 2% formaldehyde and quenched by 0.125M glycine. After lysis, cells on ice for 20 min, digested with 500 units of DpnII, then perform the proximity ligation. After reverse crosslinking, ligated DNA was purified, then sheared to 200bp and prepared as general indexed Illumina libraries. 2ug of different indexed samples were captured twice by specific biotinylated oligos, then PCR amplified streptavidin beads pulled down libraries. The NG capture-C libraries were sequenced on MiSeq V2 paired-end 100 cycles.