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Evaluating Massive Parallel Sequencing for Missing Persons Identification

NCJ Number
Date Published
February 2020
Elisa Wurmbach Ph.D.
Grant Number(s)

This is the Final Summary Overview of the findings and methodology of a research project that evaluated a proposed forensic science process for its capability to improve the identification of missing persons.


The method tested used massively parallel sequencing (MPS) in conjunction with ancestry, phenotypic and identity SNPs and STRs, as well as sequencing of the mitochondrial HV1/HV2 regions and complete genome. The focus was on the accuracy and robustness of the new markers’ correct identification, along with their ability to do this with degraded samples. Illumina’s ancestry and phenotypic SNPs were assessed on a diverse population of about 300 individuals. The project also sought to automate the complex DNA library preparation method to improve consistent results. In discussing the implications of this project for criminal justice policy and practice in the United States, the project team notes that all experiments, including forensic casework, require positive and negative controls to ensure the correct working of a kit; however, all positive controls (2800M) using the ForenSeq DNA Signature Prep kit mix B showed variable losses of typed loci. Issues with the positive control show limited validity in the runs and/or kit, as case to case comparisons could not be performed. In case of dropouts within the positive control, rules should be established for data analysis in casework as follows: 1) repeat the run; 2) disregard the group that showed dropouts, or 3) disregard the affected loci. The ForenSeq DNA Signature Prep kit Primer Mix B failed to accurately predict a missing person’s eye and/or hair color. The Illumina whole mitochondrial DNA kit did not produce successful results. The report suggests a redesign of amplicons and primer to result in better and more robust PCR reactions. 4 tables

Date Created: February 11, 2021