title: GSM6432870: Young_rep5; Mus musculus; RNA-Seq
library_construction_protocol: 5 young and 9 middle-age B6 mice harvested. Single cell suspensions from each mouse were prepared by crushing spines, sternums, humeri, tibias, femurs, and iliac crests.  Niche cells were submitted to enzymatic digest, stained, and sorted on a FACSympony S6 sorter. Hematopoietic cells pass through a ficoll gradient, stained, and sorted on a FACSAria II sorter. The cells were immediately processed as follows. Cells were counted on a Countess II automated cell counter (ThermoFisher), and 12,000 cells were loaded into one lane of a 10X Chromium microfluidic chip. Single cell capture, barcoding and library preparation were performed using the 10X Chromium version 3.1 NEXTGEM chemistry, according to the manufacturer's protocol (#CG000315). cDNA and libraries were checked for quality on Agilent 4200 Tapestation and quantified by KAPA qPCR before sequencing. each gene expression library was sequenced at 18.75% of an Illumina NovaSeq 6000 S4 flow cell lane, targeting 6,000 barcoded cells with an average sequencing depth of 75,000 reads per cell.