title: GSM8497229: B2-Caco-Mim-Gut-R2_S10; Homo sapiens; RNA-Seq
library_construction_protocol: Cell lysates were prepared from OrganoPlate® 3-lane 40 microphysiological system with flow (MPS) or static 24-well Transwells™. Following 10 minutes of incubation at room temperature, the cell lysate was flushed out by addition lysis buffer. Following 10 min of incubation at room temperature, the cell lysate was flushed out by addition of 15 uL of Enhanced 2× Lysis Buffer (201250, BioSpyder Technologies, Carlsbad, California). For Transwells, media was aspirated and 90 uL of 1× lysis buffer was added directly onto cells. After 10 min at room temperature the lysates were collected from each insert and stored at -80C until further processing.  The Templated Oligonucleotide Sequencing Assay (TempO-Seq™, BioSpyder Technologies, Carlsbad, CA) was used as an RNA sequencing technology of choice. Detailed protocols for TempO-seq are provided by the manufacturer and were previously reported  detailed elsewhere (House et al., 2017) TempO-seq libraries were prepared using the S1500+ targeted transcriptome panel (Mav et al., 2018) consisting of 2,976 transcripts according to the manufacturer's instructions.