GSM7431644: Caco-2/HT29 coculture, LPS-treated, rep 1; Homo sapiens; RNA-Seq
On day 23, cells were trypsinized to be harvested from the Transwell. Following fast thaw in 37C water bath, cell viability was checked using Thermo Fisher Countess II FL with NucBlue staining. Following cell viability counting, all samples below 70% viability were subjected to dead cell removal via annexin bead conjugation to improve quality. Single cell RNA libraries were generated using the Chromium Single Cell 3' kit (10X Genomics, CA, USA). The cells were counted and loaded onto the Chromium Controller. Loading was performed to target capture of ~3,000 GEMs per sample for downstream analysis, and samples were processed through the Chromium Controller following the standard manufacturer's specifications. AThe sequencing libraries were evaluated for quality on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA). Pooled libraries were quantified using qPCR (Applied Biosystems, Carlsbad, CA, USA) prior to loading onto an Illumina sequencing platform. The samples were sequenced at a configuration compatible with the recommended guidelines as outlined by 10X Genomics.