Effects of the Ion PGM™ Hi-Q™ sequencing chemistry on sequence data quality.
Massively parallel sequencing (MPS) offers substantial improvements over current forensic DNA typing methodologies such as increased resolution, scalability, and throughput. The Ion PGM™ is a promising MPS platform for analysis of forensic biological evidence. The system employs a sequencing-by-synthesis chemistry on a semiconductor chip that measures a pH change due to the release of hydrogen ions as nucleotides are incorporated into the growing DNA strands. However, implementation of MPS into forensic laboratories requires a robust chemistry. Ion Torrent's Hi-Q™ Sequencing Chemistry was evaluated to determine if it could improve on the quality of the generated sequence data in association with selected genetic marker targets. The whole mitochondrial genome and the HID-Ion STR 10-plex panel were sequenced on the Ion PGM™ system with the Ion PGM™ Sequencing 400 Kit and the Ion PGM™ Hi-Q™ Sequencing Kit. Concordance, coverage, strand balance, noise, and deletion ratios were assessed in evaluating the performance of the Ion PGM™ Hi-Q™ Sequencing Kit. The results indicate that reliable, accurate data are generated and that sequencing through homopolymeric regions can be improved with the use of Ion Torrent's Hi-Q™ Sequencing Chemistry. Overall, the quality of the generated sequencing data supports the potential for use of the Ion PGM™ in forensic genetic laboratories.
['Churchill|Jennifer D|JD|', 'King|Jonathan L|JL|', 'Chakraborty|Ranajit|R|', 'Budowle|Bruce|B|']
D016172:DNA Fingerprinting D000068598:Data Accuracy D053803:Forensic Genetics D005819:Genetic Markers D054629:Genome, Mitochondrial D059014:High-Throughput Nucleotide Sequencing D006801:Humans D018895:Microsatellite Repeats D044469:Racial Groups D017422:Sequence Analysis, DNA
Ion PGM™ Hi-Q™ Massively parallel sequencing (MPS) STRs Mitochondrial DNA