GSM1960257: 750N input DNA; Homo sapiens; ChIP-Seq
Flash-frozen minced tissue was thawed in cell culture medium (Gibco® Keratinocyte-SFM (1X) Catalog #17005-042) with 1% formaldehyde for 10 minutes under rotation for cross-linking, which was then quenched with 2M Glycine (final concentration of 100mM). Minced tissue was then washed twice with ice-cold PBS with Protease Inhibitor Cocktail (PIC) and resuspended in 1mL ChIP Lysis Buffer (1% SDS, 50mM Tris HCl pH 8.1, 10mM EDTA) with PIC. The suspension was ground in a dounce homogenizer 10-15 times and then passed through a 40µM filter and checked for lysis with Trypan Blue under a microscope. The nuclei were then sonicated using a probe sonicator (Branson Sonifier® Cell Disruptor 185, 4.5 Output Control, 12 cycles, 10 seconds ON/30 seconds OFF) and centrifuged at 12,000rpm for 10 minutes at 4°C. Libraries were constructed using the KAPA DNA library preparation kit (KK8230)
GEO Accession
GSM1960257