title: GSM6533600: Aging mouse AN27, scRNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: FACS sorted HSPCs including myeloid progenitors [lineage (CD3, CD4, CD8, B220, Gr1, Mac1, Ter119)-/IL-7R+/Sca1-], common lymphoid progenitors [lineage (CD3, CD4, CD8, B220, Gr1, Mac1, Ter119)-/IL7R+/Flk2+], and KLS cells [lineage (CD3, CD4, CD8, B220, Gr1, Mac1, Ter119)-/ckit+/Sca1+] from aging mice that were 30 months old. Sorted cells were washed in PBS with 0.04 % bovine serum albumin (BSA) Library was performed according to the manufacter's instructions (single cell 3' v3.1 protocol, 10x Genomics). Briefly, Bone marrow cells were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with beads, amplified by PCR and the apropiated sized fragments were selected with SPRI select reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.