Abstract

High throughput mRNA and Protein qPCR Profiling

Mikael Kubista, Professor/Founder, TATAA Biocenter AB

qPCR is developing into the most powerful platform to profile samples extracting the biologically relevant information. Major breakthrough in recent years is the development of robust preamplification methods, which allow the profiling of large number of markers starting with minute sample amounts, including single cells. In first part of my talk I will describe high throughput single cell profiling of astrocytes collected over-time from mouse brains after induced trauma. The profiling reveals how the brain responds to the injury by activating the astrocytes reacting to the trauma and inducing repair. This multiway study with expression of many genes, measured in large number of cells, over time is analyzed with powerful multivariate tools in GenEx from MultiD to identify clusters of coregulated genes, which biological functions and expression pathways are then mapped using the Ingenuity iReport. In the second part of my talk I will present high throughput protein expression data measured with qPCR using Olink Proximity extension Assays combined with preamplification.