On-chip quantification of miRNA using digital droplet PCR
Poster May 13, 2014
Q. Cai1, R.S. Wiederkehr1, B. Jones1, B. Majeed1, T. Stakenborg1, P. Fiorini1, L. Lagae1, M Tsukuda2, T. Matsuno2, I. Yamashita2
Recent advances show that miRNA has a great potential in diagnostics. Several studies demonstrate that the expression profile of miRNAs, which are short (18-22 nucleotides) but powerful regulators of a wide range of biological processes, can aid in distinguishing between healthy and diseased patients. As these regulators are detectable in peripheral blood, they can perfectly serve as blood-based biomarkers. An interesting way to profile miRNAs involves the use of lab-on-chip tools, which allow automated quantification. Therefore, it is of interest to implement miRNA release, reverse transcription (RT) and detection of miRNA into a single microfluidic device or chip. As digital droplet PCR (ddPCR) has shown to be more accurate and has an improved reproducibility compared to regular qPCR, microfluidic chips for ddPCR as well as for thermal lysis and RT were fabricated using in house silicon technology.
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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