GSM5553102: NCI-H660-ASCL1-Input; Homo sapiens; ChIP-Seq
Cell lines were grown in media supplemented with 5% FBS (Gibco #A3160701) and processed for ChIP using the Magna ChIP Kit (Millipore #17-10085) according to manufacturer's instructions with the following antibodies: ASCL1 (5 µg, Abcam#556604, EZH2 (5 μg, Active Motif #39933), H3K27Me3 (5 µg; Millipore #07-449). 10 million cells were fixed with 1% formaldehyde for 10 minutes at room temperature. Cells were collected. Chromatin was sonicated to approximately 500bp. For ChIP-seq, sequencing libraries (100 ng DNA per sample) were constructed using the KAPA library Prep Kit (Roche, Cat KK8510) with Illumina TruSeq indexes. Libraries were assessed for quality using gel electrophoresis and libraries passing quality control (e.g., no primer dimers) were quantified using the KAPA Library Quantification Kit for Illumina (Roche, Cat KK4824). Libraries were sequenced using the Illumina NextSeq 500 (75 bp single-end reads; cell lines). 