An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells

Abstract

Leakage of mitochondrial or nuclear DNA into the cytosol can occur following viral infections, radiation damage, and in some cancers. Here, we present an optimized protocol for isolating and quantifying cytosolic DNA from mammalian cells. We describe steps for collecting cytosolic fractions from cells, extracting DNA using columns, and quantifying extracted DNA using qPCR. This straightforward protocol can be completed in as little as 5 hours, and allows for identification of the source of DNA.