Influence of the electrophoresis-resequencing method on the forensic mtDNA profiling quality

Abstract 

The validation of an upgraded mtDNA profiling procedure served as a study on the electrophoresis-resequencing methods ABI377-LR5%-Sequence Navigator, ABI3100-POP4-SeqScape and ABI3100-POP6-SeqScape. The upgraded protocol was considered to be suitable for mitochondrial DNA sequencing of HV1 and HV2, with all three electrophoresis types tested. Nevertheless, the sequencing protocol had to be extended with additional primers to obtain a full double-stranded profile of samples with specific sequencing problems in the C-stretch of HV1 or HV2. The major differences between the tested electrophoresis types were observed in the detection limit and the automatic mixed base detection. But also the primer to sequencing start-distance depends on the method.

Keywords: Electrophoresis, Mitochondrial DNA, Sequencing, Validation