GSM8118479
RNA was extracted from snap-frozen tissue or cell pellets. For tissue samples, tissue was minced using a liquid nitrogen-cooled mortar and pestle before RNA extraction. RNA isolation was performed using the RNeasy Mini kit (Qiagen) including an on-column DNase I digestion. RNA quality was assessed by automated electrophoresis using the RNA 6000 Pico Kit and an Agilent 2100 BioAnalyzer (Agilent Technologies, Inc.). RNA was quantified using the Qubit RNA HS Assay Kit and a Qubit 2.0 fluorometer (Thermo Fisher Scientific). Cells were thawed, hashed with TotalSeq A anti-human hashtag antibodies (BioLegend), prepared as specified by BioLegend. Custom barcode amplification was performed by two rounds of PCR. Libraries were sequenced on the NovaSeq Illumina platform (Center for Advanced Technology, UCSF).