Characterization of NIST standard reference materials by next generation sequencing |
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| Institution: | 1. Laboratorio de Genética Molecular de Cruz Vital – Cruz Roja Ecuatoriana Quito, Ecuador;2. Laboratorio GENES Ltda, Medellin, Colombia;1. Flinders Centre for Nanoscale Science and Technology, Flinders University, Sturt Road, Bedford Park, Adelaide, South Australia, Australia;2. School of Biological Sciences, Flinders University, Sturt Road, Bedford Park, Adelaide, South Australia, Australia;1. Network of Forensic Science Institutes, Institute of Forensic Medicine, DNA Laboratory, Budapest, Hungary;2. Research Centre for Natural Sciences of the HAS, Department of Complex Systems, Budapest, Hungary;1. Institute of Legal Medicine, Faculty of Medicine, University of Cologne, Cologne, Germany;2. IPATIMUP – Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal;3. Forensic Genetics Unit, Institute of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Spain |
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| Abstract: | The National Institute of Standards and Technology (NIST) offers certified Standard Reference Materials (SRMs) for laboratories which perform DNA-based human identity testing. Developed by the Applied Genetics Group at NIST, forensic DNA SRMs are well characterized for relevant markers such as autosomal and Y-chromosomal short tandem repeats (STRs) and mitochondrial DNA (mtDNA) sequence. Characterization of the SRMs has been performed by capillary gel electrophoresis fragment-based genotyping analysis and/or Sanger type sequencing. New technologies, commonly referred to as next generation sequencing (NGS), now offer an opportunity to validate the certified DNA-based reference materials using an orthogonal method and thereby increase the confidence in the certified values of the materials. The increased sensitivity of NGS will facilitate expansion of the information content of NIST SRMs through the characterization of new features which are beyond the ability of fragment-based analysis or Sanger sequencing to detect. This work will also lead to the availability of Certified Reference Materials for laboratories wishing to implement and validate NGS technology in the field of forensics. |
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| Keywords: | Mitochondrial DNA Sanger sequencing Next generation sequencing |
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