Developmental validation of the MiSeq FGx Forensic Genomics System for Targeted Next Generation Sequencing in Forensic DNA Casework and Database Laboratories

Forensic Science International Genetics, Journal Year: 2017, Volume and Issue: 28, P. 52 - 70

Published: Jan. 27, 2017

Human DNA profiling using PCR at polymorphic short tandem repeat (STR) loci followed by capillary electrophoresis (CE) size separation and length-based allele typing has been the standard in forensic community for over 20 years. Over last decade, Next-Generation Sequencing (NGS) matured rapidly, bringing modern advantages to analysis. The MiSeq FGx™ Forensic Genomics System, comprised of ForenSeq™ Signature Prep Kit, Reagent instrument Universal Analysis Software, uses simultaneously amplify up 231 a single multiplex reaction. Targeted include Amelogenin, 27 common, autosomal STRs, 24 Y-STRs, 7 X-STRs three classes nucleotide polymorphisms (SNPs). kit includes two primer sets: 58 STRs 94 identity informative SNPs (iiSNPs) are amplified Primer Set A (DPMA; 153 loci); if laboratory chooses generate investigative leads B, amplification is targeted DPMA plus 22 phenotypic (piSNPs) 56 biogeographical ancestry (aiSNPs). High-resolution genotypes, including detection intra-STR sequence variants, semi-automatically generated with software. This system was subjected developmental validation studies according 2012 Revised SWGDAM Validation Guidelines. two-step first amplifies target STR SNP (PCR1); unique, sample-specific indexed adapters or "barcodes" attached PCR2. Approximately 1736 reactions were analyzed. Studies substrate testing (cotton swabs, FTA cards, filter paper), species from range nonhuman organisms, input sensitivity 1ng down 7.8pg, two-person human mixture genotype combinations, stability analysis partially degraded DNA, effects five commonly encountered inhibitors. Calculations repeatability reproducibility (1ng template) indicate 100.0% accuracy System calling relative CE (1260 samples), >99.1% bead array samples iiSNPs, 310 aiSNPs piSNPs), >99.0% >97.8% precision, respectively. Call rates observed all both mixes. Limitations discussed. Results described here demonstrate that meets quality assurance guidelines robust, reliable, reproducible performance on various quantities qualities.

Language: Английский

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DNA transfer in forensic science: A review

Forensic Science International Genetics, Journal Year: 2018, Volume and Issue: 38, P. 140 - 166

Published: Oct. 26, 2018

Language: Английский

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Massively parallel sequencing of forensic STRs: Considerations of the DNA commission of the International Society for Forensic Genetics (ISFG) on minimal nomenclature requirements

Forensic Science International Genetics, Journal Year: 2016, Volume and Issue: 22, P. 54 - 63

Published: Jan. 21, 2016

Language: Английский

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Recommendations of the DNA Commission of the International Society for Forensic Genetics (ISFG) on quality control of autosomal Short Tandem Repeat allele frequency databasing (STRidER)

Forensic Science International Genetics, Journal Year: 2016, Volume and Issue: 24, P. 97 - 102

Published: June 19, 2016

Language: Английский

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Massively parallel sequencing techniques for forensics: A review

Electrophoresis, Journal Year: 2018, Volume and Issue: 39(21), P. 2642 - 2654

Published: Aug. 13, 2018

DNA sequencing, starting with Sanger's chain termination method in 1977 and evolving into the next generation sequencing (NGS) techniques of today that employ massively parallel (MPS), has become essential application areas such as biotechnology, virology, medical diagnostics. Reflected by growing number articles published over last 2-3 years, these have also gained attention forensic field. This review contains a brief description first, second, third techniques, focuses on recent developments human analysis applicable Relevance to is besides standard STR-profiles, repeats can be sequenced look for polymorphisms. Furthermore, additional SNPs acquire information ancestry, paternity or phenotype. The current MPS systems are very helpful cases where only limited amount highly degraded been secured from crime scene. If enough autosomal not present, mitochondrial maternal lineage analysis. These clearly demonstrate use NGS will grow an indispensable tool science.

Language: Английский

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Microhaplotypes in forensic genetics

Forensic Science International Genetics, Journal Year: 2018, Volume and Issue: 38, P. 54 - 69

Published: Oct. 1, 2018

Language: Английский

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High‐throughput microsatellite genotyping in ecology: improved accuracy, efficiency, standardization and success with low‐quantity and degraded DNA

Molecular Ecology Resources, Journal Year: 2016, Volume and Issue: 17(3), P. 492 - 507

Published: Aug. 9, 2016

Abstract Microsatellite markers have played a major role in ecological, evolutionary and conservation research during the past 20 years. However, technical constrains related to use of capillary electrophoresis recent technological revolution that has impacted other marker types brought question continued microsatellites for certain applications. We present study improving microsatellite genotyping ecology using high‐throughput sequencing ( HTS ). This approach entails selection short suitable , PCR ‐amplified on an Illumina platform bioinformatic treatment sequence data obtain multilocus genotypes. It takes advantage fact gives direct access sequences, allowing unambiguous allele identification enabling automation process through bioinformatics. In addition, massive parallel abilities expand information content single experimental runs far beyond electrophoresis. illustrated method by brown bear samples amplified with multiplex 13 new sex marker. provided accurate individual parentage assignment resulted significant improvement success (84%) faecal degraded DNA costs reduction compared The holds vast potential success, accuracy, efficiency standardization ecological applications, especially those rely profiling low‐quantity/quality construction genetic databases. discuss give perspectives implementation light challenges encountered wildlife studies.

Language: Английский

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Characterization of genetic sequence variation of 58 STR loci in four major population groups

Forensic Science International Genetics, Journal Year: 2016, Volume and Issue: 25, P. 214 - 226

Published: Oct. 1, 2016

Language: Английский

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Recent progress, methods and perspectives in forensic epigenetics

Forensic Science International Genetics, Journal Year: 2018, Volume and Issue: 37, P. 180 - 195

Published: Aug. 17, 2018

Language: Английский

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Massively parallel sequencing of forensic STRs and SNPs using the Illumina ® ForenSeq™ DNA Signature Prep Kit on the MiSeq FGx™ Forensic Genomics System

Forensic Science International Genetics, Journal Year: 2017, Volume and Issue: 31, P. 135 - 148

Published: Sept. 8, 2017

Language: Английский

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