Simultaneous extraction and separation of rna and dna from single cells using electrophoretic techniques

Abstract

Devices and methods for preparing RNA and DNA from single cells are disclosed. In particular, the invention relates to a method of simultaneously extracting RNA and DNA from single cells and separating the nucleic acids electrophoretically. An electric field is used to lyse a single target cell, such that the plasma membrane is selectively disrupted without lysing the nuclear membrane. Cytoplasmic RNA is separated from the nucleus by performing isotachophoresis (ITP) in the presence of a sieving matrix that preferentially reduces the mobility of the nucleus. During ITP, the cytoplasmic RNA accumulates at an ITP interface between leading and trailing electrolytes and can later be extracted free of nuclear DNA. The method can be performed in a microfluidic device that fully automates all steps of the process.