GSM9159034: LSK (Lin-/Sca+/Kit+) cells - Control 2; Mus musculus; ATAC-seq
LSK cells were sorted into DMEM media without serum and spun (500g, 5 minutes, 4°C) and flowthrough discarded. Cells were washed once with 200uL cold PBS, spun down (as before) and flowthrough discarded. Pellets were resuspended in 25uL of PBS and quickly added lysis buffer (10 mM Tris-HCl pH7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% GEPAL CA-630) and centrifuged immidiately at 500 ×g 4°C for 30 seconds. Supernatant was discarded and 39ul of water immediately added, after which were also added 10uL Tagment DNA Buffer and 1uL Tagment DNA Enzyme. Cells were vortexed and incubated for 30 minuted at 37°C Immediately following transposition, samples were purified using the Qiagen MinElute PCR Purification Kit according to the manufacturer's instructions. Library construction was performed following Illumina's standard protocol for sequencing on the NovaSeq 6000 platform, including PCR amplification and addition of sequencing adapters.