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Use of standardized bioinformatics for the analysis of fungal DNA signatures applied to sample provenance
استفاده از بیوانفورماتیک استاندارد برای تجزیه و تحلیل امضاهای DNA قارچی اعمال شده برای پیشروی نمونه-2020 The use of environmental trace material to aid criminal investigations is an ongoing
field of research within forensic science. The application of environmental material thus far has focused upon a variety of
different objectives relevant to forensic biology, including sample provenance (also referred to as sample
attribution). The capability to predict the provenance or origin of an environmental DNA sample would
be an advantageous addition to the suite of investigative tools currently available. A metabarcoding
approach is often used to predict sample provenance, through the extraction and comparison of the DNA
signatures found within different environmental materials, such as the bacteria within soil or fungi
within dust. Such approaches are combined with bioinformatics workflows and statistical modelling,
often as part of large-scale study, with less emphasis on the investigation of the adaptation of these
methods to a smaller scale method for forensic use. The present work was investigating a small-scale
approach as an adaptation of a larger metabarcoding study to develop a model for global sample
provenance using fungal DNA signatures collected from dust swabs. This adaptation was to facilitate a
standardized method for consistent, reproducible sample treatment, including bioinformatics processing
and final application of resulting data to the available prediction model. To investigate this small-scale
method, 76 DNA samples were treated as anonymous test samples and analyzed using the standardized
process to demonstrate and evaluate processing and customized sequence data analysis. This testing
included samples originating from countries previously used to train the model, samples artificially
mixed to represent multiple or mixed countries, as well as outgroup samples. Positive controls were also
developed to monitor laboratory processing and bioinformatics analysis. Through this evaluation we
were able to demonstrate that the samples could be processed and analyzed in a consistent manner,
facilitated by a relatively user-friendly bioinformatic pipeline for sequence data analysis. Such
investigation into standardized analyses and application of metabarcoding data is of key importance for
the future use of applied microbiology in forensic science. Keywords: Forensic microbiology | Bioinformatics | Metabarcoding | Sample provenance |
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