COLO829 human melanoma cell line treated with DMSO
COLO829 melanoma cell line
braf signaling status
anti-MITF (Sigma, HPA003259, lot# A08324)
Sequenced DNA Library
20-50 million cells were used. Cells were trypsinized, washed, and cross-linked as described previously(Sen et al. 2008). Lysis with a modified RIPA buffer (1% NP40/Igepal, 0.5% Sodium Deoxycholate, 0.1% SDS, 100uM EDTA in PBS), nuclei were isolated and DNA was sonicated to ~150bp and overnight pulldown was performed with an antibody to MITF (Sigma HPA003259) ChIP-sequencing libraries were prepared with a modified version of NEBNext ChIP-seq library prep kit (NEB) using Ampure beads (Agencourt) for purification. Barcodes were introduced and these libraries were sequenced with 1x50bp reads on the Illumina HiSeq platform.