GSM8775463: Murine prostate DLP; Mus musculus; ATAC-seq
Cells were collected in 1 ml PBS supplemented with 3% FBS at 4 °C. Cells were centrifuged and cell pellets were resuspended in 100 μl lysis buffer (TrisHCl 10 mM, NaCl 10 mM, MgCl2 3 mM, Igepal 0.1%) and centrifuged at 500g for 25 min at 4 °C. Supernatant was carefully discarded and nuclei were resuspended in 50μl reaction buffer (Tn5 transposase 2.5μl, TD buffer 22.5μl, from Nextera DNA sample preparation kit, Illumina, and 25μl H20). The reaction was performed at 37 °C for 30 min and was stopped by adding 5μl clean up buffer (NaCl 900 mM, EDTA 300 mM). DNA was purified using the MiniElute purification kit (QIAGEN) following the manufacturer's protocol. DNA libraries were PCR amplified (Nextera DNA Sample Preparation Kit, Illumina), and size selected from 200 to 800 bp (BluePippin, Sage Sciences), following the manufacturer's recommendations.