GSM3956284
GSM3956284: ST3_H3K27ac; Homo sapiens; ChIP-Seq
Ependymoma samples were homogenized and fixed in 1% formaldehyde for 10 min at room temperature. Crosslinking was quenched with the addition of 125 mM glycine for 5 min. Samples were then lysed in SDS lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl) and sonicated with Bioruptor (Diagenode) to 200-500 bp fragments. After centrifugation, the lysates were diluted with dilution buffer (167 mM NaCl, 0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl) and incubated with 10 µg of anti-H3K27me3 antibody (9733, Cell Signaling Technology) bound to Dynal magnetic beads (Invitrogen) overnight at 4¡C. Magnetic beads were washed with low salt buffer (150 mM NaCl; 0.1% SDS; 1% Triton X-100; 1 mM EDTA; 50 mM Tris-HCl), high salt buffer (500 mM NaCl; 0.1% SDS; 1% Triton X-100; 1 mM EDTA; 50 mM Tris-HCl), LiCl buffer (150 mM LiCl; 0.5% Na deoxycholate; 0.1% SDS; 1% Nonidet P-40; 1 mM EDTA; 50 mM Tris-HCl) and TE buffer (1 mM EDTA; 10 mM Tris-HCl). ChIP DNA was eluted and recovered using Qiagen PCR purification kit. ChIP sequencing libraries were prepared according to the Illumina TruSeq protocol and were sequenced on the HiSeq 2000.
GEO
GSM3956284
GEO Accession
GSM3956284