The innovative forensic technique of DNA profiling has been acclaimed as the most important advance in forensic science since fingerprinting. Whilst there is much anecdotal information on the impact of DNA profiling on criminal investigation, prosecution and adjudication, there is little quantitative and control comparison data on the routine use of forensic DNA profiling. This study evaluates the effect of the introduction of DNA profiling in Victoria on a number of key points in the criminal justice system. The overall impact of DNA profiling was low as determined by the percentage of criminal cases which utilise DNA profiling. However, in certain classes of cases its impact was measurable, most notably in sex offences committed by “strangers”. Less than one quarter of sexual offence cases of DNA profiled resulted in a contested trial, suggesting that the focus of DNA profiling on the criminal justice system should swing to the pre-trial phase. DNA profiling was most often used in sexual offence cases and a database comparison of these cases before and after the advent of DNA profiling was examined. Whilst not statistically significant, trends indicated there were more solved cases, more guilty pleas and fewer trials after the introduction of DNA profiling, but more individuals were drawn into the investigatory process. The number of trials of sexual offences in which consent was an issue was slightly greater than previously. The increase in guilty pleas with DNA profiling was only for stranger type crime. Further research is recommended as DNA profiling becomes the cornerstone of biological forensic analysis.

The fundamental objective of conservation genetics is the identification of the basic units of conservation. Central to this objective is the reconstruction of the adaptive and evolutionary history of populations to evaluate their conservation status. Evolutionary history involves both microevolutionary and macroevolutionary processes and adaptive history is the evolution of specific characters to selective ecological processes in differential heterogeneous environments. Neutral DNA markers such as mitochondrial DNA, minisatellites and microsatellites are most often used for reconstructing history and identifying conservation units. This thesis examined three biological systems: 1) an African cichlid, 2) Canadian moose populations and 3) eastern North American wolves and coyotes to test two hypotheses. Firstly, neutral DNA markers can be used to accurately reconstruct the evolutionary history of populations. Secondly, neutral DNA markers are concordant with adaptive distinctiveness in reconstructing the adaptive history of populations. Few studies have examined these relationships. Lake Magadi tilapia showed discordant patterns between adaptive morphological, physiological and behavioural characters and genetic structure assessed with mitochondrial DNA. I propose this discordance has resulted from selection acting on mitochondrial DNA that has often been assumed to be "neutral". Neutral DNA markers accurately reflected the known history of the moose populations but discordant patterns were observed between neutral and functional loci indicating the former may not accurately reflect adaptive variation. DNA profiles of eastern wolves and coyotes showed a significant conflict in the interpretation of mtDNA and microsatellite data compared to previous genetic studies that examined wolf taxonomy. The data were consistent with the hypothesis of a North American-evolved wolf. Coyote-like mtDNA was not of coyote origin but represented divergent but related sequences of a North American wolf lineage independent of the gray wolf (C. lupus). Under this new model of eastern wolf evolution, we also identified the hybrid origin of eastern coyotes, contrary to previous interpretations, and genetically characterised different wolf "types" within Ontario. These findings could not reject the first hypothesis as neutral markers were used to reconstruct the histories of the three biological systems. However, the findings identified that it is important to ensure the neutrality of DNA markers and that samples are representative of the taxa under investigation. The findings in this thesis did not support the second hypothesis, as neutral DNA markers were not concordant with adaptive characters, i.e. morphology, physiology and functional genetic markers. / Thesis / Candidate in Philosophy

Philosophiae Doctor - PhD / DNA profiling the male specific region on the Y-chromosome is fundamental to forensic practise. Its recognised as a powerful analytical tool for investigation of sexual assault when the DNA evidence is highly admixed. Standard practises for processing sexual assault evidence include physically separate the sperm cell from the female fraction using differential extraction followed by autosomal DNA profiling. However, under specific scenarios of assault physical separation may not be possible due to the nature of the evidence. The research presented in this thesis was focused on the development and validation of the UniQTyper™ Y-10 prototype for male specific DNA profiling. The prototype which contains 10 Y-STR markers was developed and validated to deliver a rapid and cost-effective system while maintaining a forensic applicable level of performance. An allelic ladder is produced with an allele cloning approach for which an overview of the workflow and technicalities presented herein is aimed to assists an efficient bulk production process. In a second component novel sequence variation was reported across 153 sequenced alleles and submitted to Genbank. In this output the Y-STR panel was perused beyond the scope of length polymorphisms. In a proof of concept, its potential to discriminate between shared allele sizes by characterizing sequence structure variations is discussed. In a final component we generate the largest Y-STR survey across South Africa to establish reference data and to comprehensively assess the forensic genetics parameters for the UniQTyper™ Y-10.

The ability of jurors and juries to comprehend and utilise scientific evidence in Australian criminal trials has been examined. From mock jury surveys relating to DNA profiling evidence, it was determined that most respondents were able to comprehend some basic and applied statistics, although their ability was in part related to their knowledge of English and their level of education. The point at which mock jurors were prepared to convict an accused solely on the basis of DNA profiling evidence was examined and found to be low compared with the strength of DNA profiling evidence commonly presented in Australian courts. Mock jurors also demonstrated the ability to process evidence that was presented in a Bayesian framework; commencing with prior odds, introducing new information and culminating in posterior odds. From a survey of Australian forensic scientists, including fraud investigators, it was found that most practitioners' concerns could be addressed by greater pre-trial consultation between experts and legal advocates. Improved knowledge within the legal profession concerning the jargon, principles, procedures, limitations and conclusions to be drawn from different scientific disciplines, prior to presenting this evidence in court, is recommended as the means by which complex evidence can be better adduced from expert witnesses and better presented to juries in criminal trials. Finally, from interviewing actual jurors in criminal trials in the Australian Capital Territory it was determined that where jurors' expectations of scientific evidence, particularly DNA profiling evidence, are not met, high levels of juror frustration and speculation may culminate in hung juries. The adversarial setting of criminal proceedings was also found to produce an environment in which jurors felt that information that would assist them in reaching a verdict was being deliberately withheld. The ability of the jury to ask questions and the allowed nature of those questions were also examined, with the resultant recommendation that juries be given more explicit information at the commencement of trials to inform them about their rights and obligations when asking questions.

DNA profiling has ignited public interest and consequently their expectations for the capabilities of forensic criminal and science investigations. The prospect of characterising the genetic makeup of individuals or trace samples from a wide variety of depositional and post-mortem circumstances raises the question of how reliable the methods are given the potential for prolonged exposure to variation in environmental factors, i.e. temperature, pH, UV irradiation and humidity, that are known to induce damage to DNA. Thus, it is crucial to verify the validity of the DNA profiling for characterising the genetic makeup of post-mortem tissues. This project aimed to assess the reliability of sequence and microsatellite based genotyping of tissues (muscle, hair and bone) sampled from carcasses over a two year post-mortem period. This assessment investigated the impact of environment induced DNA degradation in the local geographic region that is typical of the circumstances that confront forensic practitioners in southern Australia and to utilise rigorous controls by studying animals whose time of death and burial was known and for which we had pre-decay tissue samples available. A ‘body farm’ with 12 pig carcasses on the northern Adelaide plains, ~60km north of Adelaide, which has a typical southern Australian Mediterranean climate, i.e. cold wet winters and hot dry summers. Pigs (Sus scrofa) were used as an experimental analogue for human subjects because of the logistical and ethical reasons. The pig carcasses were allocated among three treatments: four were left on the surface, four were buried at 1m depth, and four were buried at 2 m depth. These ‘burial’ conditions mimic a range of conditions encountered typically in forensic and archaeological studies. Cortical bone samples were taken from each pig carcass at one week, one month, three months, six months, one year and two years post-mortem and muscle and hair over the same sampling period for as long as those tissue types were present. A set of PCR primers to amplify two (short and a long) fragments from the hypervariable part of the mitochondrial control region (HVRI) that is used in forensic and evolutionary studies of humans and many other mammal species were developed. Also a panel of four pig microsatellite loci with fluorescent labels to facilitate automated multiplex genotyping. These loci matched as closely as possible the core motifs and allele lengths typical of the commercially available microsatellite marker kits used in Australian forensic science labs so that our experiments were as good a model as possible of the human forensic DNA technology. In this study it was possible to retrieve samples from muscle tissue up to 90 days, hair up to one year and bone at two years post-mortem. The analyses showed that the long and short HVRI region PCR fragments were only amplifiable up to 30 days from muscle tissue and that these fragments were amplifiable up to one year from hair. In contrast, in cortical bone both PCR fragments were amplifiable up to two years. The long fragment disappeared in muscle tissue completely after 30 days and in hair after six months. However, the long fragment was present in cortical bone even at two years. Overall, there was a general trend of loss of concentration of both the long and short fragments over time. Comparisons of the HVRI nucleotide sequences among tissues sampled from individual animals showed substitution changes in muscles as early as 30 days (3 out of 6 individuals) and hair at six months (1 out of 6 individuals). In contrast, in cortical bone substitutions first appeared at 365 days (1 out of 6 individuals). The most common substitution observed in all tissues types was the C-T transition, with A-G transversions observed in two episodes and C-A transversion observed in one episode. Analyses of microsatellite genotypes in muscle tissues showed high allele peaks on chromatograms up to day seven samples. However, by three months PCR was not successful from muscle tissue. While, bone tissue had lower allele peak heights compared to the muscle tissues, alleles were detectable up to six months. Allele drop out occurred for one animal (at 2 meters) in muscle tissue at the dinucleotide locus and for another animal (kept on surface) also in muscle tissue at a tetranucleotide locus. Stuttering was observed for a single animal at dinucleotide locus in muscle tissue (buried sample 2 meter depth). No stuttering or allele drop outs were seen in the bone tissue. Overall the four loci completely disappeared after 30 days in muscle tissue and after 180 days in bone tissue. In summary, analyses showed that post-mortem DNA degradation was present in all the three tissue types (muscle, hair and bone). The types of damage identified were DNA fragmentation, nucleotide substitutions and DNA loss, which resulted in a diminished frequency of successful PCR for mitochondrial and nuclear markers over time and stuttering and allele drop out in microsatellite genotyping. In addition, two nucleotide substitutions were concentrated in ‘hotspots’ that correlate with sites of elevated mutation rate in vivo. Also the frequency of successful PCR of longer nuclear and mitochondrial PCR products declined markedly more quickly than for shorter products. These changes were first observed at much shorter post mortem intervals in muscle and much longer post mortem intervals in hair and bone tissue. When considering the carcass deposition treatments, tissues that were retrieved from buried carcases showed higher levels of DNA degradation compared to tissues retrieved from carcases left on the surface. Overall, muscle tissue is a good source for DNA analysis in immediate post mortem samples, whereas hair and bone tissue are good source for DNA analysis from older samples. When comparing the microsatellite genotyping and mtDNA analyses, mtDNA is a reliable source for DNA analysis from tissue recovered from bodies that had decayed for longer post-mortem durations such as months to years, whereas microsatellite genotyping gives reliable results for tissue from shorter post mortem intervals (hours to few days). Therefore it is recommended that when analysing mtDNA sequences, cloning and sequencing PCR products can help to identify the base pair substitutions especially for tissue retrieved from longer post mortem intervals. In addition, increasing the template DNA concentrations and "neutralising" co-extracted DNA inhibitors should be considered when dealing with tissue from longer post mortem intervals. Finally, the more stringent protocols used in ancient DNA studies should be considered when dealing with tissue with much longer post mortem intervals in forensic settings. / Thesis (Ph.D.) -- University of Adelaide, School of Medical Sciences, 2009

Indiana University-Purdue University Indianapolis (IUPUI) / Forensic DNA profiling is widely used as an identification tool for associating an individual with evidence of a crime. Analysis of a DNA sample involves observation of data in the form of an electropherogram, and subsequently annotating a DNA “profile” from an individual or from the evidence. The profile obtained from the evidence can be compared to reference profiles deposited in a national DNA database, which may include the potential contributor. Following a match, a random match probability is calculated to determine how common that genotype is in the population. This is the probability of obtaining that same DNA profile by sampling from a pool of unrelated individuals. Each state has adopted various laws requiring suspects and/or offenders to submit a DNA sample for the national database (such as California’s law that all who are arrested must provide a DNA sample). These profiles can then be associated with past unsolved crimes, and remain in the database to be searched in the event of future crimes. In the case of database samples, a physical sample of the offender’s DNA must be kept on file in the laboratory indefinitely so that in the event of a database hit, the sample is able to be retested. Current methods are to collect a buccal swab or blood sample, and store the DNA extracts under strict preservation conditions, i.e. cold storage, typically -20° C. With continually increasing number of samples submitted, a burden is placed on crime labs to store these DNA extracts. A solution was required to help control the costs of properly storing the samples. FTA™ paper was created to fulfill the need for inexpensive, low maintenance, long term storage of biological samples, which makes it ideal for use with convicted offender DNA samples. FTA™ paper is a commercially produced, chemically treated paper that allows DNA to be stored at room temperature for years with no costly storage facilities or conditions. Once a sample is required for DNA testing, a small disc is removed and is to be used directly in a PCR reaction. A high quality profile is important for comparing suspect profiles to unknown or database profiles. A single difference between a suspect and evidentiary sample can lead to exclusion. Unfortunately, the DNA profile results yielded from the direct addition have been unfavorable. Thus, most crime laboratories will extract the DNA from the disc, leading to additional time and cost to analyze a reference sample. Many of the profiles from the direct addition of an FTA™ disc result in poor quality profiles, likely due to an increase in PCR inhibitors and high concentrations of DNA. Currently, standardized protocols regarding the recommended locations for removal of a sample disc from a bloodspot on an FTA™ card does not exist. This study aims to validate the optimal location by comparing DNA profiles obtained from discs removed from the center, halfway, and edge locations of a bloodspot from 50 anonymous donors. Optimal punch location was first scored on the number of failed, partial or discordant profiles. Then, profile quality was determined based on peak characteristics of the resulting DNA profiles. The results for all three disc locations were 5.3% failed amplifications, 4.2% partial amplifications, and one case of a discordant profile. Profile quality for the majority of the samples showed a high incidence of stutter and the absence of non-template adenylation. Of the three disc locations, the edge of the blood stain was ideal, due to a presumably lower concentration of DNA and likely more dilute amount of the PCR inhibitor heme. Therefore, based on the results of this study, there is a greater probability of success using a sample from the edge of a blood stain spotted in FTA™ paper than any other location of the FTA™ card.

Increasing use is being made of various types of scientific evidence in court. The general requirement for the admissibility of such evidence is relevance. Although expert evidence is considered to be opinion evidence, it is admissible if it can assist the court to decide a fact in issue; provided that it is also reliable. In South Africa, the initial wide judicial discretion to either admit or exclude unconstitutionally obtained evidence, has developed into a more narrowly defined discretion under the final Constitution. Examples of scientific evidence, namely, DNA evidence, fingerprints, psychiatric evidence, bite-mark evidence and polygraph evidence are considered and problems inherent in the presentation of such evidence in courts in various jurisdictions are highlighted. An investigation of the presentation and evaluation of evidence in both the accusatorial and inquisitorial systems seems to indicate that the adversarial procedure has a marked influence on the evaluation of evidence / Criminal & Procedural Law / LL.M. (Law)

Increasing use is being made of various types of scientific evidence in court. The general requirement for the admissibility of such evidence is relevance. Although expert evidence is considered to be opinion evidence, it is admissible if it can assist the court to decide a fact in issue; provided that it is also reliable. In South Africa, the initial wide judicial discretion to either admit or exclude unconstitutionally obtained evidence, has developed into a more narrowly defined discretion under the final Constitution. Examples of scientific evidence, namely, DNA evidence, fingerprints, psychiatric evidence, bite-mark evidence and polygraph evidence are considered and problems inherent in the presentation of such evidence in courts in various jurisdictions are highlighted. An investigation of the presentation and evaluation of evidence in both the accusatorial and inquisitorial systems seems to indicate that the adversarial procedure has a marked influence on the evaluation of evidence / Criminal and Procedural Law / LL.M. (Law)