GSM3701189: Q111SST; Mus musculus; ChIP-Seq
Compound- or vehicle-treated cells were crosslinked with 1% formaldehyde for 8 minutes and quenched with glycine for 5 minutes, lysed in 2X lysis buffer (50mM Tris-HCl pH8, 150mM NaCl, 1% Triton X-100, 0.1% Na Deoxycholate, 5mM CaCl2 and protease inhibitors) for 20 minutes on ice, and digested with 100u MNase (New England Biolabs M0247) for 10 minutes at 37°C. The MNase digestion was terminated by addition of 10mM EDTA. Chromatin was incubated with the anti-H3K4me3 antibody (Millipore 07-473, RRID: AB_1977252) overnight at 4°C, followed by incubation with Protein G beads (Invitrogen 10004D) for 2 hours at 4°C. The beads were washed with PBS (6x) and samples were eluted in EB (10mM Tris-HCl pH8, 5mM EDTA, 300mM NaCl, 0.1% SDS) supplemented with Proteinase K (New England Biolabs P8107S). SPRI beads were used for clean-up and yield was measured using Qubit Fluorimeter. Libraries were prepared using NEBNExt® Ultra™ II DNA Library Prep Kit for Illumina (New England Biolabs E7645S). Libraries were sequenced on an Illumina Hi-Seq 2000 for single-end 50bp reads.
GEO Accession
GSM3701189