Speaker: Dr Xueyi Dong (WEHI) Abstract: The lack of benchmark datasets with inbuilt ground-truth makes it challenging to compare the performance of existing long-read isoform detection and differential expression analysis workflows. Here, we present a benchmark experiment using two human lung adenocarcinoma cell lines that were each profiled in triplicate together with synthetic, spliced, spike-in RNAs (âsequinsâ). Samples were deeply sequenced on both Illumina short-read and Oxford Nanopore Technologies long-read platforms. Alongside the ground-truth available via the sequins, we created in silico mixture samples to allow performance assessment in the absence of true positives or true negatives. Our results show that StringTie2 and bambu outperformed other tools from the 6 isoform detection tools tested, DESeq2, edgeR and limma-voom were best amongst the 5 differential transcript expression tools tested and there was no clear front-runner for performing differential transcript usage analysis between the 5 tools compared, which suggests further methods development is needed for this application. About the speaker: Dr Xueyi Dong is a postdoctoral research officer in Chen lab in ACRF Cancer Biology and Stem Cells division, the Walter and Eliza Hall Institute of Medical Research (WEHI). She did her undergraduate in Zhejiang University in China, majored in biology science (2014-2018). She completed her PhD at WEHI in 2023 under the supervision of Prof. Matthew Ritchie, Dr. Charity Law and Prof. Gordon Smyth. Her current research primarily involves the analysis of spatial transcriptomics data and the investigation of RNA splicing. This event will be held online. Zoom: https://uni-sydney.zoom.us/j/84087321707