GSM7430726: OrganoidScreen; sample32; CHIR3-SAG1000; Homo sapiens; RNA-Seq
Three organoids from each condition were pooled at day 72–74, incubated in 30 U/mL papain enzyme solution (Worthington Biochemical, LS003126) and 0.4% DNase (12,500 U/mL, Worthington Biochemical, LS2007) at 37 °C for 45 minutes. Organoids were triturated with a P1000 pipette and were resus-pended in FBS followed by trituration with a P200 pipette tip. Cells were resuspended in PBS with 0.4% DNase and filtered through a 70 μm Flowmi Cell Strainer (Bel-Art, H13680-0070). The samples were centrifuged at 200 x g for 3 minutes and were resuspended in Parse Cell Buffer containing BSA. Cells were then fixed using the Parse Biosciences Cell Fixation (Parse Biosci-ences, ECF2001) kit and stored at −80°C. Barcoding and library prep was performed with the Evercode Whole Transcriptome kit (Parse Biosciences, ECW02030). Samples were prepared with v1 kits, with the exception of SAG concentration series which was prepared with v2 kits. Samples were spread across 8 sublibraries and sequenced by Admera Health on a NovaSeq S4 (Illumina).