ERX269440	SAMEA2078457	MLE_ChIP-seq	ChIP-Seq	GENOMIC	ChIP	xxx	xxx	xxx	xxx	Illumina_Genome_Analyzer_IIx	ERA223780	ERS252029	ERP003260	PRJEB4008	2014-08-12T19:16:10Z	Organism taxonomy_id="7227" taxonomy_name="Drosophila melanogaster"	Alias=E-MTAB-1705:cell_culture_1	Broker name=ArrayExpress	Description=Protocols: S2 cells were grown in Schneiders Media (Invitrogen) supplemented with 10% of heat-inactivated bovine fetal serum to a density of 1 million cells per ml. 30ml cell suspension were spun down and the cell pellet was resuspended in 1 ml of cross-linking solution (50 mM HEPES, 100 mM NaCl, 1 mM EDTA, 0.5 mM EGTA). 37% formaldehyde was added to a final concentration of 1.8%. The tube was rotated at room temperature for 15 minutes and the reaction was quenched with 2.5 M glycine at a final concentration of 125 mM for 5 minutes. The fixed cells were washed 3 times for 5 minutes each with rinse buffer 1 (10 mM HEPES pH 7.5, 10 mM EDTA, 0.5 mM EGTA, 0.25% Triton X-100) and 5 times for 5 minutes each with rinse buffer 2 (10 mM HEPES pH 7.6, 1 mM EDTA, 0.5 mM EGTA, 200 mM NaCl). Finally, the cells were washed 3 times in RIPA buffer (25 mM HEPES pH 7.5, 140 mM NaCl, 1 mM EDTA, 1% Triton-X 100, 0.1% SDS, 0.1% DOC). The cells were sonicated 30 x 20 seconds using a Branson 250 sonicator at pulse 40, intensity 3%. The chromatin should be sheared into fragments with average length of 200 bp. Samples were centrifuged at maximum speed and the supernatant was used for the subsequent steps. To reverse cross-linking, samples were incubated at 65C overnight in TE buffer, followed by a treatment with RNaseA (0.2 mg/ml) for 30 min at 37C, and with Proteinase K (0.05 mg/ml) for 2 hours at 50C. Finally, DNA was purified using Minelute columns (Qiagen). The libraries were prepared following Illumina's TruSeq ChIP sample prep kit (http://www.illumina.com/products/truseq_chip_sample_prep_kit.ilmn)	INSDC center alias=European Bioinformatics Institute (EBI)	INSDC center name=European Bioinformatics Institute (EBI)	INSDC first public=2013-10-30T17:01:19Z	INSDC last update=2018-03-08T16:29:47Z	INSDC status=public	SRA accession=ERS252029	Sample Name=ERS252029	Title=cell_culture_1	cell line=S2	cell type=embryonic cell	genotype=wild type	sex=male