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Simultaneous analysis of nuclear and mitochondrial DNA, mRNA and miRNA from backspatter from inside parts of firearms generated by shots at “triple contrast” doped ballistic models

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Abstract

When a firearm projectile hits a biological target a spray of biological material (e.g., blood and tissue fragments) can be propelled from the entrance wound back towards the firearm. This phenomenon has become known as “backspatter” and if caused by contact shots or shots from short distances traces of backspatter may reach, consolidate on, and be recovered from, the inside surfaces of the firearm. Thus, a comprehensive investigation of firearm-related crimes must not only comprise of wound ballistic assessment but also backspatter analysis, and may even take into account potential correlations between these emergences. The aim of the present study was to evaluate and expand the applicability of the “triple contrast” method by probing its compatibility with forensic analysis of nuclear and mitochondrial DNA and the simultaneous investigation of co-extracted mRNA and miRNA from backspatter collected from internal components of different types of firearms after experimental shootings. We demonstrate that “triple contrast” stained biological samples collected from the inside surfaces of firearms are amenable to forensic co-analysis of DNA and RNA and permit sequence analysis of the entire mtDNA displacement-loop, even for “low template” DNA amounts that preclude standard short tandem repeat DNA analysis. Our findings underscore the “triple contrast” method’s usefulness as a research tool in experimental forensic ballistics.

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Acknowledgments

The authors would like to thank the DFG (Deutsche Forschungsgemeinschaft) and the SNF (Swiss National Science Foundation) for funding this project. The expert technical assistance of Julia Brünig and Marion Sauer is also gratefully acknowledged.

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Authors and Affiliations

  1. Institute of Legal Medicine, University of Bonn, Stiftsplatz 12, 53111, Bonn, Germany

    Melanie Grabmüller, Jan Euteneuer, Burkhard Madea & Cornelius Courts

  2. Institute of Forensic Medicine, University of Bern, Bühlstrasse 20, 3012, Bern, Switzerland

    Christian Schyma

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  1. Melanie Grabmüller
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Correspondence to Cornelius Courts.

Electronic supplementary material

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12024_2015_9695_MOESM1_ESM.tif

Selection of a reference gene for mRNA expression data normalization using the “RefGenes” online resource (TIFF 1469 kb)

12024_2015_9695_MOESM2_ESM.tif

Imaging of PCR products resulting from two multiplex PCRs using “Mito-Mini” primer sets in “low template” samples. PCR products were separated via microfluid-gelelectrophoresis on an Agilent 2100 Bioanalyzer; Primer: primer name; bp: base pairs (TIFF 1320 kb)

Supplementary material 3 (PDF 13 kb)

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Grabmüller, M., Schyma, C., Euteneuer, J. et al. Simultaneous analysis of nuclear and mitochondrial DNA, mRNA and miRNA from backspatter from inside parts of firearms generated by shots at “triple contrast” doped ballistic models. Forensic Sci Med Pathol 11, 365–375 (2015). https://doi.org/10.1007/s12024-015-9695-3

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