GSM8860470: scRNA_sublibrary_7; Homo sapiens; RNA-Seq
Cell pellets of cultured organoids were collected and stored in Trizol for total RNA extraction using Zymo Direct-zol RNA Miniprep Kit (Zymo Research, Irvine, CA, R2063) following manufacturer's instructions. Tumor tissues were flash frozen in liquid nitrogen and ground into fine powder with the addition of Trizol, followed by RNA extraction using Zymo Direct-zol RNA Miniprep Kit. Single cell RNA seq samples were prepared using an EvercodeTM Fixation V2 kit and EvercodeTM WT V2 Kit (Parse Biosciences, Seattle, WA) following manufacturer's instructions. Briefly, organoid culture or tumor tissues were dissociated into single cells by enzyme digestion and flow sorting. The single cells were subject to a fixation procedure followed by a split-pool combinatorial barcoding protocol in 96 well format using the Evercode Kits. The barcoded cDNAs were equally dispersed into 8 indexed sub-libraries and were submitted for high throughput sequencing analysis.