title: GSM6047765: hsc pbs 1; Mus musculus; RNA-Seq
library_construction_protocol: To isolate purified LT-HSCs from the murine bone marrow, density gradient centrifugation was performed to enrich low-density mononuclear cell (LDMNCs), followed by lineage depletion of mature hematopoietic cell lineages using rat anti-mouse biotin-conjugated lineage antibodies and Biotin Binder Dynabeads (Life Technologies). LT-HSCs were then isolated from lineage-negative bone marrow cells by fluorescence activated cell sorting. Single LT-HSC cells were subsequently captured on a small sized IFC using the Fluidigm C1 system. Briefly, cells were washed, re-suspended in PBS supplemented with C1 suspension buffer in a 4:1 ratio and 400 cells/µl were loaded onto the chip. After cell capture, each position on the chip was imaged and only single cells were included in the downstream library preparation and analysis. Sequencing libraries were produced with the Illumina Nextera XT kit according to the adopted Fluidigm protocol. Bulk samples, i.e. 100 cells pooled together and then processed as with the same protocol. Bulk samples in 5week data are:  "AS-117588-LR-16803", "AS-117747-LR-16804", "AS-117900-LR-16805", "AS-118051-LR-16806", "AS-118210-LR-16807", "AS-118357-LR-16808". In the 20 week data, the bulk samples are: "AS-172797-LR-27156", "AS-172545-LR-27143" and "AS-172630-LR-27144".