GSM9752619: Dpp9-CSPL; Homo sapiens; RNA-Seq
The femur is surgically isolated and single-cell suspension of bone marrow cells are obtained through crushing the femur with a mortar and pestle and suspended in FACS buffer (PBS +2% FBS). Human CD45+ cells expressing CD34 and negative for lineage markers (CD3, CD14, CD16, CD19, CD20, CD56) and negative for mouse CD45 and mouse C-KIT are sorted on Bigfoot Spectral cell sorter. Control and DPP9KO cells are hashtagged separately and then pooled. 10,000 cells were encapsulated into droplets using 10x Chromium GEM technology. Libraries were prepared in house using the Chromium Next GEM Single Cell 3′ Reagent Kits v.3.1 with 3' feature barcode kit (10x Genomics). scRNA-seq libraries were sequenced using the Nova-Seq system (Illumina).