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                            Library & Multimedia
 Systematic evaluation of the Precision ID GlobalFilerTM NGS STR panel v2 using single-source samples of various quantity and quality and mixed DNA samples
NCJ Number
              308391
Journal

*Forensic Science International: Genetics* Volume: 69 Dated: March 2024 Pages: 102995
Author(s)
          Vishakha Sharma;
                        Elisa Wurmbach
Date Published

2024
Annotation
              This paper provides a systematic evaluation the Precision ID GlobalFilerTM NGS STR panel v2 using single-source samples of various quantity and quality and mixed DNA samples.
Abstract
              This study evaluated in 24 experimental runs the Precision ID GlobalFilerTM NGS STR panel v2 from ThermoFisher, which targets 31 autosomal STRs, amelogenin, and three Y-markers (one STR, SRY, and Yindel). Single-source samples were used in 18 experimental runs, for systematic evaluation. These included assessing library preparation benchmark conditions, limited DNA input, as well as testing repeatability, number of samples per run, and degraded DNA samples. Full profiles were consistently obtained from as little as 50 pg DNA input. Using the optional recovery PCR method improved outcomes for samples with low DNA input. Full profiles were also obtained from severely degraded DNA samples with degradation indices (DI) of > 60. In addition, six experimental runs were performed testing various two-person mixtures with mixture ratios ranging from 1:20 to 20:1. Major and minor contributors were distinguishable by their read counts (coverage), because less DNA input yielded lower read counts, analogous to the traditional CE technology, where less DNA produces lower peak heights. Mixture ratios of approximately 1:1 were indistinguishable, while a greater imbalance, i.e., higher mixture ratios, made the mixture more distinguishable between major and minor contributors. Based on this information, the highest success rate of correctly deconvoluted four-allelic loci was from mixtures with 1:3 ratios. At higher mixture ratios, the drop-out rate of the minor contributor increased, reducing the number of four-allelic loci. Massively parallel sequencing (MPS) techniques were developed approximately 15 years ago. Meanwhile, several MPS kits for forensic identification, phenotypic information, ancestry, and mitochondrial DNA analysis have been developed. Sequencing short tandem repeats (STRs) has certain advantages over the currently used length-based genotyping methods. MPS is more discriminative and includes the possibility of testing high numbers of targets, different types of markers, as well as the use of smaller amplicons. (Published Abstract Provided)
      
  
          Additional Details
Grant Number(s)
        2018-DU-BX-0166


  
  
        
  Agencies
        NIJ-Sponsored


  
  
        
  Publication Type
        Research (Applied/Empirical)


  
  
        
  Award
        Evaluation of Massively Parallel Sequencing Technology for Routine Forensic Casework
    
  
  
Date Published: January 1, 2024
    
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        DNA typing



  
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