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Development of Reference Sample DNA Profiling for Databases Using Next Generation Sequencing Technologies

NCJ Number
Date Published
February 2015
112 pages
Findings and methodology are reported for a project whose goals were to develop and evaluate massively parallel sequencing (MPS) systems that can type reference samples for the entire mtGenome and a large battery of autonomic, Y-chromosome, and X-chromosome STRs and human identity SNPs in a single multiplex.

Most forensic laboratories focus only on the control region of the mtGenome and, more specifically, hyper variable regions I and II (HVI and HVII) for database construction, database queries, and direct and indirect sample comparisons; however, MPS may make it practical to sequence the entire mtGenome in a rapid and facile manner. The number of markers will provide more robust associations and significantly reduce candidate lists. It is likely that as more kinship associations result in solving crime, there will be motivation to further use familial searching with MPS profiling. Overall, project goals were met. Large multiplex systems were developed and obtained, followed by testing for typing reference samples. STRs 9autosomal, X chromosome and Y chromosome, and identity SNPs could be typed simultaneously. SNPs were also typed in their own multiplex. Whole mtGenomes could be sequenced with relative ease. The data support that reliable results can be obtained. In order to facilitate analyses, software was developed. STRait Razor (v1.0 and v2.0) for STR typing and mitSAVE for haplotype alignment/nomenclature have been created and are freely available. The protocols described in the final report and published in the scientific literature should enable novel users to perform MPS in their respective laboratories. 19 tables, 12 figures, and 133 references

Date Published: February 1, 2015