EVALUATION OF THE PERFORMANCE OF THE BETA VERSION OF THE FORENSEQ DNA SIGNATURE PREP KIT ON THE MISEQ FGx FORENSIC GENOMICS SYSTEM

Abstract Massively Parallel Sequencing (MPS) are innovative approaches of analysis of genetic markers, supported by the forensic community. The Illumina Beta Version of the ForenSeq DNA Signature Prep Kit is a high-throughput, multiplexing kit allowing simultaneous, high resolution sequencing of 153 identity informative markers (including 27 autosomal STRs, 7 X-chromosomal, 24 Y-chromosomal haplotype markers and 94 SNPs) plus 56 biogeographical ancestry and 22 phenotypic informative SNPs. Implementation of MPS technology in forensic casework laboratories needs comprehensive, internal validation according to SWGDAM guidelines. The aim of this study was to determine with ForenSeq DNA Signature Prep Kit, method's precision/accuracy, sensitivity as well as possibility of analysis of two persons mixtures. Analysis of challenging casework samples coming from various tissues were performed and compared to results obtained with capillary electrophoresis (CE). In order to evaluate the quality and reliability of the acquired results, informative metrics of STR and SNP markers including depth of coverage (DoC), allele coverage ratio (ACR), and sequence coverage ratios (SCRs) were assessed.