13.04.2024 | Short Communication
NSPlex: an efficient method to analyze non-specific peaks amplified using commercial STR kits
verfasst von:
Yukinobu Kutsuwada, Sho Tomotake, Hidetoshi Tsuda, Kazuhisa Watanabe, Ayumi Matsumoto, Sadahiko Iwamoto, Natsuko Mizuno
Erschienen in:
International Journal of Legal Medicine
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Abstract
Commercial short tandem repeat (STR) kits exclusively contain human-specific primers; however, various non-human organisms with high homology to the STR kit's primer sequences can cause cross-reactivity. Owing to the proprietary nature of the primers in STR kits, the origins and sequences of most non-specific peaks (NSPs) remain unclear. Such NSPs can complicate data interpretation between the casework and reference samples; thus, we developed “NSPlex”, an efficient method to discover the biological origins of NSPs. We used leftover STR kit amplicons after capillary electrophoresis and performed advanced bioinformatics analyses using next-generation sequencing followed by BLAST nucleotide searches. Using our method, we could successfully identify NSP generated from PCR amplicons of a sample mixture of human DNA and DNA extracted from matcha powder (finely ground powder of green tea leaves and previously known as a potential source of NSP). Our results showed our method is efficient for NSP analysis without the need for the primer information as in commercial STR kits.