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The presented study aims at gaining a better understanding of factors affecting the variability of trace DNA profiles obtained from biological material recovered from firearms handled in gun-related crimes. For this purpose, skin contact traces resulting from four realistic alternative handling scenarios were recovered from different surfaces of two types of firearms and the corresponding trace profile characteristics (DNA yield, relative profile contribution for known and unknown contributors) were compared. Observed differences between handling conditions, firearms and surface types, handling individuals and individual deposits emphasize the complexity of trace DNA profile composition expected to be recovered from firearms after realistic handling scenarios. The obtained data will serve as useful information for forensic scientists providing expert opinion on alternative propositions at activity level in the context of gun-related crimes.

In molecular ballistics, the molecular genetic analysis of biological material is integrated with the biophysical wounding events and trace investigations of terminal ballistics. It aims to support the objective, evidence-based reconstruction of gun-related crimes. Of central interest is the analysis of backspatter traces: biological material propelled back from the entry site of a bullet after impact on a biological target. This study is a first approach to systematically investigate possible correlations or patterns between the amount of recoverable DNA (yield) from backspatter traces found on and inside fired guns and the shooting distance. After firing shots from five distances (0, 5, 10, 15, 30 cm) with both a pistol and revolver in triplicates, no clear correlations or patterns can be observed, due to considerable variation between measurements from identical triplicates and a lack of linear relation between distance and yield, e.g. yields from even 30 cm shots will sometimes exceed those of contact shots. Wound channel evaluation was performed as well and distinct differences between contact shots and distance shots could be observed, but not between any distant shots.

In recent years, forensic mRNA profiling has increasingly been used to identify the origin of human body fluids. By now, several laboratories have implemented mRNA profiling and also use it in criminal casework. In 2018 the FoRNAP (Forensic RNA Profiling) group was established among a number of these laboratories with the aim of sharing experiences, discussing optimization potential, identifying challenges and suggesting solutions with regards to mRNA profiling and casework. To compare mRNA profiling methods and results a collaborative exercise was organized within the FoRNAP group. Seven laboratories from four countries received 16 stains, comprising six pure body fluid / tissue stains and ten mock casework samples. The laboratories were asked to analyze the provided stains with their in-house method (PCR/CE or MPS) and markers of choice. Five laboratories used a DNA/RNA co-extraction strategy. Overall, up to 11 mRNA markers per body fluid were analyzed. We found that mRNA profiling using different extraction and analysis methods as well as different multiplexes can be applied to casework-like samples. In general, high input samples were typed with high accuracy by all laboratories, regardless of the method used. Irrespective of the analysis strategy, samples of low input or mixed stains were more challenging to analyze and interpret since, alike to DNA profiling, a higher number of markers dropped out and/or additional unexpected markers not consistent with the cell type in question were detected. It could be shown that a plethora of different but valid analysis and interpretation strategies exist and are successfully applied in the Forensic Genetics community. Nevertheless, efforts aiming at optimizing and harmonizing interpretation approaches in order to achieve a higher consistency between laboratories might be desirable in the future. The simultaneous extraction of DNA alongside RNA showed to be an effective approach to identify not only the body fluid present but also to identify the donor(s) of the stain. This allows investigators to gain valuable information about the origin of crime scene samples and the course of events in a crime case.

Firearms are the most relevant items of evidence in gun-related crimes, likely bearing various traces facilitating an objective reconstruction of the crime. Trace DNA recovered from firearm surfaces might help to identify individual(s) having handled the firearm and thereby possibly to link the firearm and the corresponding shooter, however, the interpretation of DNA traces on handled items can be challenging and requires a detailed understanding of various factors impacting DNA prevalence, transfer, persistence and recovery. Herein, we aimed at improving our understanding of factors affecting the variability of trace DNA characteristics recovered from firearms handled in gun-related crimes: Skin contact traces were recovered from various outer surfaces of two types of firearms handled in four realistic, casework-relevant handling scenarios and the corresponding trace characteristics (DNA yield, number of contributors, relative profile contribution for known and unknown contributors, LRs) were compared. Trace DNA characteristics differed distinctly between handling conditions, firearm and surface types as well as handling individuals and intraindividual deposits emphasizing the variability and complexity of trace DNA profile composition expected to be recovered from firearms after realistic handling scenarios. The obtained results can provide useful insights for forensic experts evaluating alternative activity level propositions in gun-related crimes.

Since DNA from touched items and surfaces ("touch DNA") can successfully and reliably be analyzed, the question as to how a particular DNA containing sample came to be from where it was recovered is of increasing forensic interest and expert witnesses in court are increasingly challenged to assess for instance whether an incriminatory DNA sample matching to a suspect could have been transferred to the crime scene in an innocent manner and to guess at the probability of such an occurrence. The latter however will frequently entail expressing a subjective probability i.e. simply making a best guess from experience. There is, to the present date, an extensive and complex body of literature on primary, secondary, tertiary and even higher order DNA transfer, its possibility, plausibility, dependency on an array of variables and factors and vast numbers of permutations thereof. However, from our point of view there is a lack of systematic data on DNA transfer with existing research widely varying in quality and relevance. Our aim was, starting from a comprehensive survey of the status quo and appreciating its increasing importance, to in the first part of our review raise consciousness towards the underestimated and insufficiently accounted for complexity of DNA transfer and thus appendant research of forensic scientists serving as expert witnesses in court but also acting in the role of a journal referee to point them to areas of criticism when reviewing a manuscript on DNA transfer. In the second part, we present propositions how to systematize and integrate future research efforts concerning DNA transfer. Also, we present a searchable database providing an extensive overview of the current state of knowledge on DNA transfer, intended to facilitate the identification of relevant studies adding knowledge to a specific question and thus help forensic experts to base their opinion on a broader, more complete and more reproducible selection of studies.