Multiplex Y-STRs analysis using the ion torrent personal genome machine (PGM).

Area of Science:

• Forensic genetics
• Genomic technologies
• Molecular biology

Background:

• Massively parallel sequencing (MPS) offers advanced capabilities for analyzing multiple DNA samples and targeted regions simultaneously.
• The routine application of MPS in forensic genetics is rapidly developing.
• Y-STR (Y-chromosome Short Tandem Repeat) analysis is crucial for forensic investigations.

Purpose of the Study:

• To evaluate the effectiveness of MPS technology for forensic Y-STR analysis.
• To develop and test a multiplex assay for 13 Y-STR loci using MPS.

Main Methods:

• A multiplex assay targeting 13 Y-STR loci was designed for MPS.
• The assay was used to amplify DNA from 42 unrelated males.
• Amplicons were sequenced using the Ion Torrent Personal Genome Machine (PGM) system.

Main Results:

• All 13 Y-STR loci were successfully detected, with DYS389 II showing a 1.8% failure rate due to amplicon size.
• New alleles were identified across several loci (DYS389 II, DYS390, DYS437, DYS448, DYS635, DYS438) due to variations in repeat composition and nucleotide substitutions.
• An inconsistency in DYS390 allele calls compared to capillary electrophoresis (CE) was observed, attributed to upstream deletions.

Conclusions:

• MPS Y-STR typing successfully identified new alleles and variations, offering greater genetic detail.
• This technology demonstrates potential for increased discriminatory power in forensic identification.
• MPS is a promising tool for advancing forensic Y-STR analysis.