Abstract

Taking Expression Profiling to New Dimensions

Mikael Kubista, Professor/Founder, TATAA Biocenter AB

qPCR, dPCR and NGS are developing into the most powerful platforms to profile samples and extract the biologically relevant information. Major breakthrough in recent years is the development of robust preamplification methods and library preparations allow for the profiling of large number of markers starting with minute sample amounts, including single cells. In the first part of my talk I will describe high throughput single cell profiling with focus on the experimental workflow and analysis of data, exemplifying with the profiling of individual astrocytes from mouse brains reflecting response to trauma. I will also present multiway studies, where expression of many genes in different types of samples/cells are measured over time and how most valuable information about the molecular processes involved and expression pathways can be extracted with powerful multivariate tools such as dynamic PCA available in GenEx. In the second part of my talk I will present single cell multianalyte profiling, showing how DNA, mRNA, microRNA, lncRNA, and protein can be measured in the same cell. I will present high throughput protein profiling using qPCR as platform, and I will present qPCR tomography to measure intracellular mRNA and microRNA profiles. Finally, I will present new molecular tools to test the quality of mRNA in clinical samples for profiling experiments and how to validate the diagnostic assays.