Project description:The goal of this study was to evaluate the transcriptional response of 4 human duodenal enteroid lines on monolayers to norovirus infections (GII.4). Enteroids were plated as monolayers in Intesticult (Stem Cell Technologies) proliferation medium. After 1 day of cell growth as a monolayer, the proliferation medium was changed with differentiation medium for 5 days. Five-day-differentiated monolayers were washed and were either mock-infected or inoculated with human norovirus supplemented with 500 μM glycochenodeoxycholic acid (GCDCA), for 1 to 2 h at 37°C. Total RNA was extracted using the Qiagen RNeasy kit and paired-end Illumina sequencing was performed.
Project description:The mutation status of TP53 was analyzed by next generation sequencing using the Illumina TruSight Tumor (TST) 15 assay (Illumina) at the Molecular Pathology Center of the Jewish General Hospital (Montreal, QC, Canada). The CAP-compliant clinically validated TST15 assay was performed as per supplier's instruction with 10 ng DNA input. Libraries were sequenced on a MiSeq with a v3 flow cell (Illumina). Data was analyzed with a clinically validated pipeline (NextGene, SoftGenetics) and variants were annotated in Geneticist Assistant (SoftGenetics). The IARC TP53 database version R18 was consulted for the molecular interpretation of TP53 variants (www.p53.iarc.fr).
