engleski

Sequencing workflow for the whole mitochondrial genomes from Croatian reference samples

Recognizing the huge potential of next generation sequencing (NGS) technology, Forensic Science Centre “Ivan Vučetić”, located in Zagreb, established the first Croatian NGS forensic laboratory at the beginning of 2016. The primary goal of our laboratory is gradual introduction of NGS methods for DNA analysis into forensic casework, by means of development, validation and implementation of forensically relevant protocols. Analysis of mitochondrial DNA (mtDNA) in today’s routine forensic practice is restricted to hypervariable sites residing in control region of the molecule. However, developing whole mtDNA sequencing is important because it considerably increases the power of discrimination when reporting the results of forensic analyses, and enables maximal recovery of mtDNA from highly degraded samples. Given that the emergence of NGS technology potentiated sequencing of the whole mitochondrial genome (16, 569 bp) at acceptable time-cost ratio, we selected this particular analysis to be the first step in the process of introducing NGS into our work. Here we present the established NGS workflow that resulted in high-quality whole mtDNA sequences obtained from reference buccal swab samples. This method will be further employed for creating a national database of whole mtDNA sequences, and thus will make foundation for the use of mtDNA analysis in forensic casework.

NGS ; Whole mtDNA sequencing ; Reference samples ; Croatian population ; Forensics

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