title: GSM7504758: PBMCs, control YTg, replicate 2; Mus musculus; RNA-Seq
library_construction_protocol: PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque (TBD Science, Tianjin, China), and mononuclear sections were collected and washed three times by PBS. Then the PBMCs were processed for scRNA-seq. Library  was performed according to the manufacter's instructions (single cell 3' Reagent Kits v3.1 (Dual Index)  protocol, 10x Genomics). Briefly, GCs 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 Silane DynaBeads, amplified  by PCR and the apropiated sized fragments were selected with SPRIselect  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.