Improvements, factors, and influences on DNA recovery from firearms.

Touch DNA recovery from firearms can be central to many criminal investigations, yet the generation of DNA profiles from these items remains poor. Currently in Australia, published casework data highlights extremely poor DNA success from samples recovered from firearms. Only between 5% and 25% of samples result in useful DNA data and therefore increasing the success of DNA recovered from firearms is highly important but has not yet been explored in-depth. This study focused on increasing the recovery of DNA from ten firearm components that were held for 15 s. Multiple recovery methods were used, and the resulting genetic data compared. DNA evidence may be deliberately removed from firearms after discharge to hamper forensic investigations, therefore this study examined the effect of wiping down the components or handling them with gloves. A standard double swab and rinse swab recovery method resulted in an average of 73% cellular recovery. A cumulative swab process had the highest average recovery at 86%, although it was found that increasing the DNA yield led to an increase in mixture complexity. Wiping over the components was observed to remove on average 69% of cellular material, compared with 33% when handed with gloves. However, the size and texture of the components affected the efficiency of cellular material removal. The results from this study allow for prioritisation of areas to sample on firearms, as well as suggesting techniques that can be applied for the optimum process of cellular recovery and subsequent generation of STR DNA data.

Persistence of touch DNA on commonly encountered substrates in different storage conditions.

The persistence of touch DNA deposited after realistic handling of items typically encountered in forensic investigations has been the subject of few studies. Understanding the long-term persistence of touch DNA on different substrates in varying conditions can be central to the effective triage of samples for further processing. As the time between an alleged incident and collection of evidence may vary from a few days to years after an alleged event, this study assessed three different common substrates for the persistence of touch DNA over a time span up to 9 months. These substrates included fabric, steel, and rubber, each of which were handled in a way to imitate what may happen during a criminal act. The three substrates were exposed to two different environments for up to 9 months: inside a dark cupboard with no traffic to act as a control and an outside semi-exposed environment. Ten replicates from each of the 3 substrates were tested at 5 time points to create 300 samples. All samples were processed using a standard operating workflow to provide genotype data after exposure to different environments. It was found that the fabric samples produced informative STR profiles (defined here as 12 or more alleles) up to the 9 month timepoint for either environment. The rubber and steel substrates for the inside condition produced informative STR profiles up to the 9 month timepoint, but only generated informative STR profiles for the outside condition up to 3 and 6 months, respectively. These data add to our understanding of the external factors that affect DNA persistence.

The influences of dusty environments on the STR typing success of post-detonation touch DNA samples.

As the use of improvised explosive devices (IEDs) in a broad spectrum of offences continues, it is vital that research is performed to assess the capabilities of the forensic DNA profiling technology currently available to provide information as to potential perpetrators. This work investigates some of the most important gaps in our understanding surrounding the poor success rates in DNA profiling obtained through the sampling of touch DNA on post-detonation IED samples. It has been previously suggested that the use of Diamond™ Nucleic Acid Dye may fix cells to a surface, therefore reducing the effect of an experimental process to remove or damage those cells. This was found not to be the case for samples undergoing a detonation as there was no difference in the resultant post-detonation profiles between the stained samples, stained prior to detonation, and unstained samples. The comparison of data from previously performed research, within an enclosed explosives chamber, to real-world outdoor detonation events in a rural and dusty environment was investigated. It was found that there was a significant difference between the environments for the aluminium but not for the battery or electrical tape substrates indicating that environment has the potential to influence STR success through the introduction of PCR inhibitors; humic acid within rural natural dust was introduced here. No difference was observed in cell loss due to the detonation between environments and the dirt within the PCR was higher in the 'outdoor' samples. The effect on cellular retention and damage due to the sample's distance from the charge has been thoroughly investigated through incremental 100 mm exposure. Distance from the charge was found to affect every metric analysed; these being the cell loss from samples, the number of alleles amplified in resultant direct PCR profiles, and the total RFU of the subsequent profiles. These data outline the importance of this work allowing results to be assessed and triage decisions be made accordingly. The analysis of wood, PVC pipe, a mobile phone with rubber buttons, a SIM card, and a circuit board showed that none of these samples at 400 mm from the charge caused substrate specific PCR inhibition. On-site collection teams do not need to triage collection based on these sample types as there was no significant difference observed in their ability to return DNA profiling data. Surface area and inhibitor presence are key variables to consider when determining STR processing workflow for post-detonation samples as for samples with larger surface areas within the outdoor environment PCR post-extraction is preferential to direct PCR.

Analysis of rapid HIT application to touch DNA samples.

Rapid DNA technology is being utilized for reference profiles worldwide. There is also strong data in the literature to support its use for high-template DNA sources, the same is not true for low-template sources, such as touch DNA; this is a requirement before wider implementation to forensic casework is considered. We report on the Rapid HIT Intel cartridge's ability to facilitate successful amplification of touch DNA to obtain profiles from template deposited on items commonly encountered in forensic casework. Eight items were touched in ten replicates- two were tapelifted, three swabbed, and three directly inserted. Significance was observed in the alleles amplified and RFU with respect to sample type. Three samples performed well: cable tie, fabric, and matchstick. As two of these were directly inserted, this should be considered for any sample small enough. Placement of highly absorbent substrates into the cartridge is not advised as it can cause a lysate-pull error. Heterozygote loci often presented as homozygous (32%-78% loci per profile); this was influenced by substrate type and profile RFU. Loci with larger masses exhibited higher false homozygosity also. Comparison of the donor's profile analyzed was performed against previous datasets analyzing touch DNA through standard workflow, including manual DNA extraction, PCR, and CE separation. These data show that for all substrates, except for a fabric swatch, standard processing is preferential to Rapid HIT analysis. In its current form, rapid DNA technology is not fit for the routine analysis of touch DNA samples in forensic casework.

DNA deposited in whole thumbprints: A reproducibility study.

Touch DNA is pivotal in forensic science therefore understanding the mechanisms and variations of deposition and composition of genetic material in touched deposits is essential. Shedder status is still poorly understood, and the consistency and cohesiveness of research is less developed compared to other transfer and persistence considerations. In this study, the inter- and intra-variations between shedder categories and individuals were investigated by use of a nucleic acid binding dye. Ten volunteers deposited 30 thumbprints under two different time points post handwashing: 15 after a period of 15 min post handwashing (defined) and 15 after a period of at least 60 min post handwashing (undefined). Thumbprints were made on glass slides, marked with a grid of 55 squares, then the marks were stained with Diamond Dye and the cells that fluoresced in each square (7500 total) counted to determine the total number of cells. Shedders were less consistent in cellular deposition when thumbprints were made in the defined condition compared to waiting at least 60 min. Heavy shedders consistently generated informative profiles (defined here as 12 or more alleles); this occurred 73% of the time in the defined condition and 87% in the undefined. Intermediate shedders produced informative profiles, which occurred 50% of the time in the defined and 80% in the undefined condition. Light shedders consistently produced uninformative profiles with only 33% being considered uploadable to a DNA database for the defined condition and 27% informative for the undefined condition.