title: GSM9378679: Control, whole bone; Mus musculus; RNA-Seq
library_name: GSM9378679
library_construction_protocol: scRNA-Seq was conducted in femoral bone samples from 5-month-old male LepR-Cre;Hes1∆/∆ and sex-matched controls. Intact femoral bones were excised, the marrow flushed, and bones digested with collagenase, dispace and DNase, incubated with ACK lysis buffer to remove red blood cells followed by immune and erythroid cell depletion by double negative (CD45−;TER119−) selection by fluorescence activated cell sorting. Barcoded libraries were prepared using a ChromiumX instrument and a Chromium Single Cell GEM-X Universal 3' gene expression v4 4-plex library and a GEM-X OCM3' Chip v4 4-plex kit. cDNA amplification and library generation were performed according to manufacturer's protocol. cDNAs from a single cell had the same barcode, allowing the sequencing reads to be mapped back to the cell of origin. Libraries were validated for length and adapter dimer removal using the Agilent TapeStation D1000 High Sensitivity assay, quantified and sequenced as one sample pool on an AVITI Platform using 2 x 7.5 high-output flow cell. Target read depth of 50,000 transcriptome reads per cell was achieved in each sample.