The mitochondrial single-stranded DNA-binding protein (mtSSB) regulates the 
function of the mitochondrial DNA (mtDNA) replisome. In vitro, mtSSB stimulates 
the activity of enzymatic components of the replisome, namely mtDNA helicase and 
DNA polymerase gamma (Pol γ). We have demonstrated that the stimulatory 
properties of mtSSB result from its ability to organize the single-stranded DNA 
template in a specific manner. Here we present methods employing electron 
microscopy and enzymatic assays to characterize and classify the mtSSB-DNA 
complexes and their effects on the activity of Pol γ.