title: GSM8370021: MLO_WT0307d60; Homo sapiens; RNA-Seq
library_construction_protocol: Three to four organoids were used for each dissociation reaction, following manufacturer's protocol. In brief, 50 units of papain enzyme (Worthington) was activated with 1 mM L-cysteine and 0.5 μM EDTA in Earle's Balanced Salt Solution (EBSS) (Thermo Fisher), and DNase I (Worthington) added. hMLOs were finely chopped with a scalpel and then resuspended in papain enzyme mix. The slurry was incubated at 37°C, 5% CO2 for 45 min on a rotator, and then triturated. Cells were spun down at 300xg for 5 min, and washed twice in EBSS containing ovalbumin (Worthington) and BSA. Library preparation for RNA-seq was performed by Novogene AIT Singapore, using a directional mRNA library prep kit (NEB) with polyA enrichment. Paired-end 150 nt sequencing was performed on the Illumina NovaSeq, with sequencing depth of at least 30 million read pairs per sample. Datasets were analyzed using the nf-core/RNA-Seq pipeline version 1.4.2 .