GSM9617795: Thalamocortical Assembloid, KOLF2, clone 1, Week 6, Non-Targeting Control; Homo sapiens; RNA-Seq
Organoids were dried by removing all media then flash frozen and stored at -80°C in preparation for single-nuclei RNA-sequencing. Nuclei were extracted from frozen organoids using the 10X Chromium Nuclei Isolation Kit using a 4-minute lysis time. Nuclei were washed twice using the included buffer, counted using a hemacytometer, then resuspended in buffer to achieve a target capture of 10,000 nuclei. During counting, nuclei were also imaged to ensure healthy morphology and lack of debris. For single-cell samples, organoid tissue was dissociated with papain plus DNase (Worthington) and processed via FACS. Samples were captured using the 10X Genomics Chromium Single Cell 3' Reagent Kit v3.1 or v4 and 10X Chromium System v3.1 or the PIPseq T2 3' Single Cell RNA Kit v4.0. All samples were assessed for quality control purposes at both the cDNA and library stages using the Agilent 4200 TapeStation System. Single-nuclei and single-cell libraries were sequenced using the Illumina NovaSeq 6000 or NovaSeq X platform.