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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When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.
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