Performance Comparison of Digital microRNA Profiling Technologies Applied on Human Breast Cancer Cell Lines
AuthorKnutsen, Erik; Fiskaa, Tonje; Ursvik, Anita; Jørgensen, Tor Erik; Perander, maria; Lund, Eiliv; Seternes, Ole Morten; Johansen, Steinar
MicroRNA profiling represents an important first-step in deducting individual RNA-based regulatory function in a cell, tissue, or at a specific developmental stage. Currently there are several different platforms to choose from in order to make the initial miRNA profiles. In this study we investigate recently developed digital microRNA high-throughput technologies. Four different platforms were compared including next generation SOLiD ligation sequencing and Illumina HiSeq sequencing, hybridization-based NanoString nCounter, and miRCURY locked nucleic acid RT-qPCR. For all four technologies, full microRNA profiles were generated from human cell lines that represent noninvasive and invasive tumorigenic breast cancer. This study reports the correlation between platforms, as well as a more extensive analysis of the accuracy and sensitivity of data generated when using different platforms and important consideration when verifying results by the use of additional technologies. We found all the platforms to be highly capable for microRNA analysis. Furthermore, the two NGS platforms and RT-qPCR all have equally high sensitivity, and the fold change accuracy is independent of individual miRNA concentration for NGS and RT-qPCR. Based on these findings we propose new guidelines and considerations when performing microRNA profiling.
PublisherPublic Library of Science (PLoS)
CitationPLoS ONE (2013), vol. 8(10): e75813.
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