Illumina HiSeq 3000 sequencing; Single cell RNA sequencing of LSK (lineage negative, Sca-1+, c-Kit+) cells isolated by fluorescence-activated cell sorting from the bone marrow of control young C57Bl/6Jax mice
Single cell RNA sequencing of LSK (lineage negative, Sca-1+, c-Kit+) cells isolated by fluorescence-activated cell sorting from the bone marrow of control young C57Bl/6Jax mice
20,000 LSK (CD4/CD8a/CD11b/GR1/B220/TER119- lineage negative, Sca-1+, c-Kit +) cells were pooled and sorted from two pooled 3-months-old female C57Bl/6Jax mice into 0.4% BSA in PBS. n/a SiC RNA-sequencing was performed on the 10X Genomics platform using the Single Cell 3â€™ Reagent Kit V3 (10x Genomics) following the manufacturer's instructions. Cells were loaded according to the manufacturer's instructions aiming for a targeted cell recovery of 8,000 cells. The quality of the obtained cDNA library upon adapter ligation and index PCR (13 cycles), was assessed by Bioanalyzer fragment analysis (HS DNA Kit, Agilent).