title: GSM2335678: mut_unt_runx1; Mus musculus; ChIP-Seq
library_construction_protocol: Frozen cell pellets (10e7 cells) were cross-linked by formaldehyde (1% final concentration) and washed by PBS. Cells were lysed and chromatin was Covaris E-220 Sonicator (Covaris). Chromatin fragments were incubated with protein G and A Sepharose beads (GE Healthcare) for two hours at 4oC to eliminate non-specific binding. Unbound chromatin fragments were separated from the beads and incubated with RUNX1 (Abcam; ab23980) antibody at 4oC overnight in IP buffer (10mM Tris-HCl pH 7.5, 2mM EDTA, 90mM NaCl, 0.1% Triton X-100, 0.1% Deoxycholate, 0.1% SDS). After incubation, IPs were washed twice with ChIP wash buffer (20mM Tris-HCl pH 8.0, 2mM EDTA, 150mM NaCl, 1% Triton X-100, 0.1% SDS) and once with final ChIP wash buffer (20 mM Tris-HCl pH 8.0, 2 mM EDTA, 500 mM NaCl, 1% Triton X-100, 0.1% SDS). IPs were eluted in elution buffer (1% SDS, 100mM Sodium Bicarbonate) and incubated at 68oC for 2 hours to reverse crosslinks. DNA fragments were stripped from histones and purified using QIAquick PCR Purification Kit (Qiagen) and subjected to 8 cycles of paired-end indexing PCR. Illumina sequencing  libraries were generated as previously described. Libraries were PCR amplified and sequenced on the Illumina NextSeq 500 platform following the manufacturer’s protocols (Illumina, Hayward CA.) Genomic DNA/RNA was extracted using the Qiagen AllPrep Mini Kit according to the manufacturer's instructions.
attributes.tag: GEO Accession
attributes.value: GSM2335678