GSM5962417: Hu218Iris; Homo sapiens; RNA-Seq
Dissected tissues were flash frozen on dry ice and stored at -80 deg C until processing. Frozen tissue samples were homogenized in a dounce homogenizer with NP-40 based lysis buffer and strained to remove debris. The nuclei were isolated by centrifugation and flow cytometry sorting with DAPI staining. The sorted nuclei were resuspended in 0.04% non-acetylated BSA in PBS to solutions of ~1000 cells/uL and loaded into Chromium Single Cell Chips for a targeted recovery of 10,000 cells.    Single nuclei libraries were generated with Chromium 3' v3 or v3.1 platform (10X Genomics, Pleasanton, CA) following the manufacturer's protocol. Single nuclei were partitioned into Gel beads in EMulsion (GEMs), where nuclei were lysed and mRNA was barcoded in reverse transcription. The cDNA was amplified, sheared, and attached to a 5' adaptor and sample index barcode. On average, 7,000-10,000 single nuclei were loaded on each channel and 4,000-6,000 cells were recovered. Libraries were sequenced on Illumina HiSeq 2500 or NovaSeq. Chromium 3' v3 or v3.1 platform (10X Genomics, Pleasanton, CA) 