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                            Library & Multimedia
 Sequencing the hypervariable regions of human mitochondrial DNA using massively parallel sequencing: Improved methods for DNA samples encountered in forensic testing
NCJ Number
              256082
Journal

*Journal of Legal Medicine* Volume: 17 Issue: 2 Dated: March 2015 Pages: 123-127
Author(s)
          C. Davis;
                        D. Peters;
                        D. Warshauer;
                        J. King;
                        B. Budowle
Date Published

March 2015
Length
              5 pages
Annotation
              In this study, 18 buccal swabs, three different tissue samples from five individuals, and four bone samples from casework were sequenced at hypervariable regions I and II, using Sanger-type sequencing (STS) and massively parallel sequencing (MPS).
Abstract
              Mitochondrial DNA testing is a useful tool in the analysis of forensic biological evidence. In cases where nuclear DNA is damaged or limited in quantity, the higher copy number of mitochondrial genomes available in a sample can provide information about the source of a sample. Currently, Sanger-type sequencing (STS) is the primary method to develop mitochondrial DNA profiles. This method is laborious and time consuming. Massively parallel sequencing (MPS) can increase the amount of information obtained from mitochondrial DNA samples while improving turnaround time by decreasing the numbers of manipulations and more so by exploiting high throughput analyses to obtain interpretable results. In the current study, sample enrichment for STS and MPS was PCR-based. Library preparation for MPS was performed using Nextera® XT DNA Sample Preparation Kit and sequencing was performed on the MiSeqTM (Illumina, Inc.). MPS yielded full concordance of base calls with STS results, and the newer methodology was able to resolve length heteroplasmy in homopolymeric regions. This study demonstrates short amplicon MPS of mitochondrial DNA is feasible, can provide information not possible with STS, and lays the groundwork for development of a whole genome sequencing strategy for degraded samples. (publisher abstract modified)
      
  
          Additional Details
Grant Number(s)
        2012-DN-BX-K033


  
  
        
  Agencies
        NIJ-Sponsored


  
  
        
  Award
        Development of Reference Sample DNA Profiling for Databases Using Next Generation Sequencing Technologies
    
  
  
Date Published: March 1, 2015
    
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      PatentsRelated Topics
        
                DNA (Deoxyribonucleic Acid)
        Forensic sciences
        Evidence analysis and processing
        Compromised DNA
        Evidence analysis



  
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