title: GSM9012197: Breast cancer organoid T_BB475; Homo sapiens; RNA-Seq
library_construction_protocol: The supernatant was discarded and 2 mL of TrypLE were used to disrupt the hydrogel structure. The suspension was then mixed by pipetting and checked under the microscope every 2 minutes until it reached a single-cell suspension state. The suspension was immediately diluted in 40 mL of cold PBS and filtered through a 35 µm sterile cell filter. The flowthrough was centrifugated at 300g for 5 minutes at 4 °C. Supernatant was discarded, the pellet was resuspended in 50 µL of cold PBS-0.02% BSA and subsequently sequenced. Library was performed according to manufacturer's instructions (Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1, 10X Genomics). Cells were resuspended in a master mix to create GEMs, and retrotranscription from polyA RNA tails was performed, resulting in a barcoded cDNA. cDNA was amplified by PCR, cleaned and used for library construction.