GSM9228307: Apollo Study sample 10 t2h NMP; Homo sapiens; RNA-Seq
Kidney tissues were cut in small pieces and isolated by nuclei isolation buffer (Nuclei EZ lysis buffer, N3408-200ML, Sigma-Aldrich) supplemented with Protease and RNase inhibitors. The nuclei were stained with DAPI (D9542-10MG, Sigma-Aldrich) and fluorescence-activated nuclei sorting was performed via Sony SH800 to enrich the nuclei. 18.000 to 20.000 nuclei per sample were loaded into the chromium controller (PN-120223, 10x Genomics) on a Chromium Next GEM Chip G (PN-2000177, 10x Genomics) and processed following the manufacturer's original protocol.Quality control for the constructed library was performed using a High Sensitivity DNA Assay (5067-4626, Agilent technologies). Libraries were sequenced on NovaSeq6000 (Illumina) targeting 50,000 reads per cell. The library construction was performed using the Chromium Next GEM Single Cell 3ʹ GEM, Library & Gel Bead Kit v3.1 (PN-1000121, 10x Genomics) and Dual Index Kit TT Set A (PN-1000215, 10x Genomics).