The primary purpose of this study was to evaluate the use of massively parallel sequencing (MPS) in conjunction with ancestry, phenotypic, and identity SNPs and STRs, along with sequencing of the mitochondrial HV1/HV2 regions and complete genome, for the purpose of improving the identification of missing persons.
The research project was conducted by the New York City Office of Chief Medical Examiner (NYCOCME). The following forensic MPS kits were assessed: 1) the ForenSeq DNA Signature Prepkit Primer MixB, for nuclear DNA, and 2) Promega PowerSeq 46GY System Prototype, for mitochondrial DNA. In discussing the implications of project findings for criminal justice policy and practice in the United States, this report notes that all experiments, including forensic casework, require positive and negative controls to ensure the correct working of a kit; however, all positive controls using the ForenSeq DNA Signature Prep kit Mix B, showed variable losses of typed loci. Issues with the positive control indicate limited validity in the run and/or kit as case-to-case comparisons cannot be performed. In case of dropouts, within the positive control, rules should be established for data analysis in casework as follows: repeat the run; disregard the group that showed dropouts; or just disregard the affected loci. The Illumina whole mitochondrial DNA kit did not produce successful results, since the longer of the two PCR reactions covering the whole genome did not result in an amplicon in any of the 31 reactions, including the positive control. This report suggests a redesign of amplicons and primers to result in better and more robust PCR reactions. 4 tables
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