Sequencing of invasive PDO8 organoids cultured in normoxia
Invasive organoids were collected from collagen and RNA was extracted using the Quick-RNA Miniprep Plus kit (Zymo Research). Libraries were prepared using the TruSeq stranded RNA-sequencing library kit (Illumina), with 30–100 ng of RNA from each condition. For every normoxic and hypoxic invasive pair, the starting RNA input was matched for library preparation. The libraries were sequenced on the Illumina NovaSeq 6000 platform with paired-end reads and coverage of 40–60 million reads per sample. 