title: GSM8637952: DN1, Del_TE, scRNAseq; Mus musculus; RNA-Seq
library_name: GSM8637952
library_construction_protocol: Bone marrow progenitors attached to OP9-DL1 stromal cells were washed three times with PBS before scraping in HBH buffer and passing through a 70uM mesh filter to minimize OP9-DL1 stromal cell contamination. Cells were then washed with PBS and fixed in 4% paraformaldehyde for 15 minutes on ice. After fixation, cells were pelleted at 500xg for 3 minutes at 4°C before resuspending in 1mL of PBSR [PBS, pH 7.4, 1% BSA, 1% SUPERase-In RNase Inhibitor (ThermoFisher Scientific), and 1% 10mM DTT], pelleting again at 500xg for 5 minutes at 4°C and resuspending in PBSR. Thawed nuclei were permeabilized with 0.2% TritonX-100 (in nuclei wash buffer) for 3 min on ice, and briefly sonicated (Diagenode, 12 sec on low power mode) to reduce nuclei clumping. The nuclei were then washed once with nuclei wash buffer and filtered through 1 ml Flowmi cell strainer. Filtered nuclei were spun down at 500×g for 5 min and resuspended in nuclei wash buffer. Libraries were prepared and sequenced as described in (Cao et al., 2019). scRNA-seq (sciRNA-seq3)
identifiers.namespace: GEO
identifiers.id: GSM8637952_r1