title: GSM2171418: HEK293_input DNA; Homo sapiens; ChIP-Seq
library_construction_protocol: The ChIP DNA was collected using the SimpleChIP™ Enzymatic Chromatin IP Kit (Cell Signaling Technology #9005), according to the manufacturer’s protocol. Briefly, HEK293 cells were collected from 15 cm plates and treated with 1 % formaldehyde (cross-linking) for 10 min and 125 mM Glycine for another 5 min (quenching) at room temperature. After washing three times with 1X PBS, cells were lysed in buffer A for 10 min on ice, then washed once in buffer B. Cells were re-suspended in buffer B, then treated with MNase (2000 gel units per 100 ml reaction) at 37 ˚C for 30 min, before the digestion was stopped by adding EDTA (10 μl 0.5 mM EDTA per 100 μl reaction). Chromatin extracts were harvested following complete lysis of nuclei by sonication. Libraries were constructed using Wafergen’s PrepX Complete ILMN 32i DNA Library Kit (cat number 400076) with an input of 5ng where available using the Apollo 324