ORCID Profile
0009-0000-3869-2725
Current Organisation
Australian Federal Police
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Publisher: Wiley
Date: 02-2023
Abstract: Formalin‐fixed tissues provide the medical and forensic communities with alternative and often last resort sources of DNA for identification or diagnostic purposes. The DNA in these s les can be highly degraded and chemically damaged, making downstream genotyping using short tandem repeats (STRs) challenging. Therefore, the use of alternative genetic markers, methods that pre‐ lify the low amount of good quality DNA present, or methods that repair the damaged DNA template may provide more probative genetic information. This study investigated whether whole genome lification (WGA) and DNA repair could improve STR typing of formaldehyde‐damaged (FD) tissues from embalmed cadavers. Additionally, comparative genotyping success using bi‐allelic markers, including INDELs and SNPs, was explored. Calculated random match probabilities (RMPs) using traditional STRs, INDEL markers, and two next generation sequencing (NGS) panels were compared across all s les. Overall, results showed that neither WGA nor DNA repair substantially improved STR success rates from formalin‐fixed tissue s les. However, when DNA from FD s les was genotyped using INDEL and SNP‐based panels, the RMP of each s le was markedly lower than the RMPs calculated from partial STR profiles. Therefore, the results of this study suggest that rather than attempting to improve the quantity and quality of severely damaged and degraded DNA prior to STR typing, a more productive approach may be to target smaller licons to provide more discriminatory DNA identifications. Furthermore, an NGS panel with less loci may yield better results when examining FD s les, due to more optimized chemistries that result in greater allelic balance and licon coverage.
Publisher: Springer Science and Business Media LLC
Date: 2013
Publisher: Springer Science and Business Media LLC
Date: 2012
Publisher: Future Science Ltd
Date: 03-2014
DOI: 10.2144/000114148
Abstract: Forensic analysis of genetic material is often limited by the quantity and quality of DNA available for examination. Stochastic effects associated with low amounts of starting template can lead to a reduction in the quality of the result, making interpretation difficult. This paper presents an lification method to copy target DNA in a linear fashion prior to short tandem repeat (STR) analysis to increase the available starting template without introducing the lification bias seen in other methods used to increase the sensitivity of PCR. Results show that implementing the pre- PCR procedure allows for greater allele recovery in multiplex STR analysis compared with s les that were not subjected to prior processing.
Publisher: Future Science Ltd
Date: 10-2018
Abstract: In forensic casework, compromised s les often possess limited or degraded nuclear DNA, rendering mitochondrial DNA a more feasible option for forensic DNA analyses. The emergence of massively parallel sequencing (MPS) has enabled the recovery of extensive sequence information from very low quantities of DNA. We have developed a multiplex PCR method that lifies the complete mitochondrial genome in a range of forensically relevant s les including single cells, cremated remains, bone, maggot and hairs isolated from dust bunnies. Following library preparation, MPS yields complete or nearly complete mitochondrial genome coverage for all s les. To confirm concordance between s le types and between sequencing platforms, we compared sequencing results from hair and buccal swabs from two references. Low initial DNA input into the multiplex PCR allows for conservation of precious DNA while MPS maximizes recovery of genetic information.
Publisher: Springer Science and Business Media LLC
Date: 09-10-2017
DOI: 10.1007/S00414-017-1698-Z
Abstract: Historically, rootless hair shaft s les submitted to a forensic laboratory for DNA analysis are reserved for mitochondrial DNA (mtDNA) analysis due to the presence of highly degraded as well as insufficient amounts of nuclear DNA. Although mtDNA has been very successful in obtaining results from rootless hair, this system has its limitations, namely, it is a lineage marker that cannot differentiate between maternally related genotypes. Given the high incidence of hairs as forensic evidence, there is a need for the use of a nuclear DNA test system capable of producing reliable results for hair shaft forensic evidence. This study reports the utilization of an enhanced DNA extraction methodology for hairs, in combination with a recently developed novel, nuclear DNA typing assay, InnoTyper® 21, to improve the success rate for obtaining informative results from highly compromised, degraded, and trace forensic s les such as rootless hair shafts. The InnoTyper 21 kit is a small licon retrotransposon marker typing system compatible with currently used capillary electrophoresis platforms. This system contains 20 Alu element markers, ranging in size from 60 to 125 bp, making the assay highly sensitive for extremely degraded forensic s les and thus enabling recovery of nuclear DNA profiles from s les that would otherwise require mtDNA sequencing. A subset of s les was also tested with the GlobalFiler kit with less success due to the larger licon sizes in comparison with InnoTyper 21. Results were variable but very promising, with approximately 40% of the total number of hairs tested producing interpretable nuclear DNA profiles with InnoTyper 21. These results demonstrate the ability of the utilized methodologies to produce nuclear DNA results with high statistical power from rootless hair shafts.
No related grants have been discovered for Kelly Grisedale.