GSM9056665: Vehide; Mus musculus; RNA-Seq
To control the final number of cells captured, approximately 30,000 cells per sample were loaded onto 10X Chromium Single Cell Platform (10X Genomics) at a concentration of 1,000 cells per μl (Single Cell 3′ library and Gel Bead Kit v.3) as described in the manufacturer's protocol Library was performed according to the manufacter's instructions (single cell 3'v3protocol, 10x Genomics). Briefly, GCs were resuspended in the master mix and loadedtogether 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 GEMwas retrotranscripted to cDNA, which contains an llumina R1 primer sequence, UniqueMolecular ldentifier (UMl) and the 10x Barcode. The pooled barcoded cDNA was thencleaned up with Silane DynaBeads, amplified by PCR and the apropiated sizedfragments were selected with SPRlselect reagent for subsequent library construction.During the library construction llumina R2 primer sequence, paired-end constructs withP5 and P7 sequences and a sample index were added.