Quantification of Global MicroRNA Abundance by Selective Isotachophoresis

摘要

We here present and demonstrate a novel techniquenbased on isotachophoresis (ITP) for the quantification ofnglobal microRNA (miRNA) abundance in total RNA. Wenleverage the selectivity of ITP to concentrate miRNA andnexclude longer RNA molecules from the focused zone. Wendesigned a novel ITP strategy where we initially establishnthree contiguous zones of sieving polymer, electrolyte, andndenaturant concentrations. This allows for successivenpreconcentration, selection, and detection of miRNA. Wenoptimized chemistry in each zone for high sensitivity andnexquisite selectivity for miRNA. This technique allows fornthe measurement of the total miRNA content in a samplenand its comparison between different cell types andntissues. We demonstrated and validated the efficacy of thisntechnique by comparing global miRNA abundance innsubconfluent and confluent cell cultures.