Combined Extraction Method for Mitochondrial DNA and Proteins from Hair for Human Identification

As submitted by the proposer: Hair is an evidentiary sample that typically does not provide sufficient nuclear DNA for forensic analysis. Thus, state-of-the-art forensic analyses for hair samples includes subjective microscopic evaluation and mitochondrial DNA (mtDNA) analysis, which can result in imprecise and inadequate conclusions. However, recent research suggests protein sequence variation in hair can be exploited for human identification purposes. Unfortunately, forensic hair research with limited sample amounts has been performed with a focus on either DNA or protein in isolation, thus limiting the forensic potential of hair samples. The Battelle team propose a fundamental/basic research 12-month program to develop and demonstrate a method that can be used to obtain both whole mt-genome haplotype and proteomic profiles from a single limited hair sample. The method will provide much larger discriminatory power than the current state-of-the-art without disrupting current forensic practices. The goal of the proposed project will be accomplished through the team’s genomics and proteomics expertise, experience, and facilities. The Battelle team will conduct logical tests and evaluations of already-conceptualized and partially tested workflows for the recovery and analysis of mtDNA and proteins from various hair samples. Protein and mtDNA will be analyzed using protein and DNA sequencing technology (mass spectrometry and massively parallel sequencing, respectively). The workflow will be developed and verified across two independent laboratories (Battelle and the University of California, Davis) using hair samples collected from a cohort of diverse individuals and multiple sources (head, pubic, and limb) to rigorously test the workflow across a range of hair types. The proposed research program will provide a foundation for the implementation of an mtDNA-compatible proteomic workflow. Once fully implemented and validated, the Battelle team expects the method could be used in forensic crime labs for routine human identification from hair samples. The resulting data from the developed workflow will dramatically and objectively increase the probability of exclusion above what the traditional molecular biological analysis of hair offers. Data generated by our program will minimally be disseminated via publishing in at least one peer-reviewed scientific journal, and at least one presentation at a forensic conference in addition to informal webinars, newsletters, and discussions with the forensic community. Note: This project contains a research and/or development component, as defined in applicable law, and complies with Part 200 Uniform Requirements - 2 CFR 200.210(a)(14). ca/ncf