title: High-throughput Sequencing of Trace Quantities of Soil Provides Reproducible and Discriminative Fungal DNA Profiles.
abstract: High-throughput sequencing (HTS) offers improved resolution between forensic soil samples by characterizing individual taxa present; however, the heterogeneous distribution of taxa in soils, and limited quantity of material available, may hinder the reliability of HTS in casework. Using HTS of the internal transcribed spacer, we examined the effect of soil mass (50, 150, and 250 mg) on fungal DNA profiles, focusing on reproducibility and discriminatory power between close proximity soils, and samples with similar textural classification. The results show that reduced soil mass had no significant effect on sample differentiation and that 150 mg soil provides the most reproducible DNA profiles across different soil types. In addition, Ascomycota was identified as a robust fungal target for forensic intelligence as this phylum was detected consistently across all samples regardless of sample quantity. Overall, this study highlights the value of trace quantities of soil for use in forensic casework.
authors.affiliation: Australian Centre for Ancient DNA, School of Biological Sciences, University of Adelaide, Darling Building, North Terrace Campus, Adelaide, SA, 5005, Australia.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Young
authors.affiliation: Australian Centre for Ancient DNA, School of Biological Sciences, University of Adelaide, Darling Building, North Terrace Campus, Adelaide, SA, 5005, Australia.
authors.forename: Laura S
authors.initials: LS
authors.lastname: Weyrich
authors.affiliation: Australian Centre for Ancient DNA, School of Biological Sciences, University of Adelaide, Darling Building, North Terrace Campus, Adelaide, SA, 5005, Australia.
authors.forename: Alan
authors.initials: A
authors.lastname: Cooper
mesh_terms: D016172:DNA Fingerprinting; D004271:DNA, Fungal / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005658:Fungi / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012988:Soil Microbiology
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer
keywords: DNA profiling; forensic science; fungi; high-throughput sequencing; internal transcribed spacer; soil