Increasing the Efficiency of Mitochondrial DNA processing of Reference Samples

Mitochondrial DNA (mtDNA) analysis in forensic testing is especially useful for human skeletal remains and hair samples. When a profile is generated for unidentified human remains or hair samples, comparison to a reference sample is critical to the case. The following steps are involved in the processing of family reference samples for mtDNA: DNA extraction, HV1 and HV2 amplification, cycle sequencing, electrophoresis, and analysis. Some procedures require DNA quantification and normalization. Analysis of mtDNA is expensive and time-consuming. A direct lysis and amplification method was previously shown to eliminate the need for DNA extraction, quantification, and normalization of reference samples. This study was performed to further develop, optimize, and validate its use for future implementation in routine casework of reference samples. The results have shown that quality mtDNA sequencing data can be obtained using a direct amplification method from blood and buccal samples from a variety of collection devices.