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                            Library & Multimedia
 Recovery of mtDNA From Unfired Metallic Ammunition Components With an Assessment of Sequence Profile Quality and DNA Damage Through MPS Analysis
NCJ Number
              252692

Author(s)
          Mitchell M. Holland, Rachel M. Bonds, Charity A. Holland, Jennifer A. McElhoe
Date Published

March 2019
Length
              11 pages
Annotation
              This study compared two methods for recovering mitochondrial (mt) DNA from the surface of unfired copper projectiles, brass casings, and aluminum casings, finding that using a cotton swab moistened with 0.5M EDTA was the favored approach, especially when the metallic surface was etched.
Abstract
              Recovery of suitable amounts of quality DNA from copper and brass surfaces, like those encountered in ammunition, has been a challenge for the forensic community. The ability of copper ions to rapidly facilitate oxidative damage leading to fragmentation of DNA significantly reduces the pool of templates for PCR amplification. In the current study, degradation was significantly higher for DNA samples recovered from copper and brass surfaces, when compared to aluminum. Massively parallel sequencing (MPS) of the control region, using the PowerSeq¢â CRM Nested System kit and the Illumina MiSeq instrument, produced full haplotypes for aluminum samples regardless of the method used to deposit or collect DNA; on the other hand, less than 60 percent of the copper and brass samples produced partial or full profile information. Touch DNA collected from copper and brass samples produced higher rates of partial or full MPS profile information (¡­88-96 percent), and collection with 0.5M EDTA produced better results than when collection was performed with water; average of ¡­70  versus ¡­47 percent. Although MPS data was not impacted by noise in the sequencing process, a higher than expected rate of noise was observed, potentially due to an increase in low-level damage lesions. Noise patterns were strikingly different when compared to control data, suggesting that noisy sites may be predictable when testing samples with high levels of oxidative damage. Library preparation was a poor predictor of MPS data quality, since a large percentage of reads did not align with the reference genome. This may impact the number of samples that can be run when a deep-coverage MPS approach is being considered for analysis of mtDNA heteroplasmy. Overall, when applying an MPS approach to the analysis of mtDNA recovered from ammunition, results are expected from touch DNA, will be limited for copper and brass components when the DNA is exposed to an aqueous environment, and DNA degradation will be accelerated when DNA comes in contact with copper or brass surfaces. Practitioners should consider collecting DNA from metallic surfaces with 0.5M EDTA, since this will maximize yield and mitigate degradation. The results of this study directly impact MPS analysis of minor mtDNA sequence variants from metallic surfaces, and are particularly relevant to forensic investigations.(publisher abstract modified)
      
  
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        Assessing the Effects of DNA Damage on Next Generation mtDNA Sequencing
    
  
  
Date Published: March 1, 2019
    
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