<p>Forensic scientists demanded a portable device that allowed individualization to facilitate their DNA analysis workflow in the field. A MinION (Oxford Nanopore Technology) is one of the potential choices for this essential task. The small-sized device came with a simple library preparation and provided sequence-based DNA analysis. In this study, blood-impregnated FTA cards were directly amplified with IDseek<sup>®</sup> OmniSTR™ Global Autosomal STR (Nimagen) and prepared for the Miseq and MinION platforms. The STRspy reference database was modified to be compatible with Nimagen kit and showed 96.55% STR genotype accuracy compared to Miseq’s data. The sensitivity test indicated that STR genotyping using Oxford Nanopore Technology can be performed in less than 0.25 ng of DNA input. The combination of a direct FTA card, Nimagen’s reverse complementary PCR, and a high-resolution MinION sequencing device demonstrated interesting performance for working in the field. The findings of this study will be added to the other previous studies in which the aim was to implement Oxford Nanopore Technology for forensic DNA analysis.</p>

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Assessing direct FTA cards for human identification using Oxford nanopore sequencing

  • Tikumphorn Sathirapatya,
  • Wikanda Worrapitirungsi,
  • Poonyapat Sukawutthiya,
  • Hasnee Noh,
  • Kornkiat Vongpaisarnsin

摘要

Forensic scientists demanded a portable device that allowed individualization to facilitate their DNA analysis workflow in the field. A MinION (Oxford Nanopore Technology) is one of the potential choices for this essential task. The small-sized device came with a simple library preparation and provided sequence-based DNA analysis. In this study, blood-impregnated FTA cards were directly amplified with IDseek® OmniSTR™ Global Autosomal STR (Nimagen) and prepared for the Miseq and MinION platforms. The STRspy reference database was modified to be compatible with Nimagen kit and showed 96.55% STR genotype accuracy compared to Miseq’s data. The sensitivity test indicated that STR genotyping using Oxford Nanopore Technology can be performed in less than 0.25 ng of DNA input. The combination of a direct FTA card, Nimagen’s reverse complementary PCR, and a high-resolution MinION sequencing device demonstrated interesting performance for working in the field. The findings of this study will be added to the other previous studies in which the aim was to implement Oxford Nanopore Technology for forensic DNA analysis.