Length bias-free extraction of cytoplasmic RNA from single cells by electrical lysis and electrophoresis

We report numerical and experimental investigations on the dynamics of RNA from single cells in the electrophoretic extraction with a focused electric field. Fluorescence microscopy uncovered two distinct kinetics of soluble and mitochondrial RNA molecules. Soluble RNA molecules show repeatable dynamics with a mean extraction time constant of 150 ms, while the mitochondria show heterogeneous dynamics with a mean extraction time of 3.8 s. Further, numerical simulations revealed that electrophoretic extraction of cytoplasmic RNA is free of the length dependence because of the uniform electrophoretic mobility of RNA molecules. Our results showed the strength of electrophoretic extraction for accurate analysis of subcellular gene expressions.