title: GSM6203824: wild type gfp+ cells; Mus musculus; RNA-Seq
library_construction_protocol: Individual plaques were microdissected from the brachiocephaic artery, miced and digested for 1 hour in Liberase plus actinomycin D to inhibit transcription during the digestion process. Samples were filtered and resuspended in PBS plsu 0.04% UltraPure non-acetylated BSA, counted, and submitted for sequecinging targeting 2,000 cells. Remaining cells were sorted on a FACS Aria fusion cell sorter and GFP+ and tomato+ cells were collected for each libary, again targeting 2,000 cells Library was performed according to the manufacter's instructions (single cell 3' v2 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 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 and paired-end constructs with P5 and P7 sequences were added.