title: GSM1184314: Med1 ChIP DP; Mus musculus; ChIP-Seq
library_construction_protocol: 50 million sorted CD4+ CD8+ DP thymocytes were fixed in 10% FCS DMEM media containing 1% formaldehyde. The cells were lysed and sonicated in [20 mM Tris-HCl, pH8, 150 mM NaCl, 2mM EDTA, 0.1% SDS, 1% Triton X, protease inhibitors]. Lysates were cleared by centrifugation and Med1-DNA complexes were captured with 5 μg anti-Med1 antibody (Bethyl, #A300-793A) and 25 μl Dynabeads. The captured DNA was eluted from the beads with a mixture of RnaseA, proteinaseK in [TE, 1% SDS] for 2 hours at 37° and then 16 hours at 65°. DNA was purified by phenol-chloroform extraction and 10ng was used with a Illumina ChIP-seq sample preparation according to the manufacturer instructions.