The quantification of DNA using PCR technology has come under scrutiny recently as MIQE guidelines suggest that qPCR information is inaccurate without following standards such as efficiency, or the accurate doubling of DNA. DNA quantification is important in research, forensic investigations, and medical diagnostics so the technique accuracy is vital. This project examined the impact of varying this efficiency value. Additionally, this research examined the steps necessary to optimize a custom mitochondrial qPCR assay.