Gene Expression Profiling: qPCR Toolkit for Quality Control
Poster Apr 05, 2012
Švec D., Jacobsson J., Sjöback R., Kubista M.
Gene expression profiling is an exceedingly important tool in functional genomics research and in drug discovery. The general trend in the field is towards higher throughput of less complex samples. Nowadays studies are appearing where expression of tens of genes is measured in hundreds of individual single cells, to be able to characterize cell subtypes and its roles. To identify subpopulations of astrocytes, to define them in molecular terms, and to determine the biological progress of astrocyte maturation on single cell level we developed and optimized protocol for high-throughput gene expression profiling using qPCR together with a system of quality control: ValidPrimeTM - control for genomic DNA background, Interplate calibrator - tool, which is used to compensate for inter-run variation in qPCR and Universal Spike (RNA/DNA), which can be used to detect presence of inhibition in samples of any living species or monitor losses and yield of isolation, or check the integrity of mRNA during sample processing, storage and transporation.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE