NCJ Number
240800
Journal
Forensic Science International: Genetics Volume: 6 Issue: 6 Dated: December 2012 Pages: 810-818
Date Published
December 2012
Length
9 pages
Annotation
This study explored the applicability of second generation sequencing (SGS) to sequence multiplexed forensic STR amplicons, both in a single contributor sample and in multiple-person mixtures with different ratios.
Abstract
The authors compared the results of a commercial STR profiling kit (Applied Biosystems AmpFlSTR() Profiler Plus()), analyzed both with capillary electrophoresis and with Roche GS FLX sequencing. An easy to use open-source software pipeline is provided, chaining together the different steps needed to start the analysis from a GS FLX FASTA file, resulting in a FASTA file containing the called and quantified alleles present in the data. Sequencing of multiplexed STR amplicons using Roche GS FLX titanium technology is technically feasible but the technology is not ideal for this purpose. The fraction of full length reads is small and the homopolymer sequencing error rate is high. The pipeline compresses the homopolymers to a single base to avoid false results caused by these homopolymers. The qualitative and quantitative results from the SGS STR analysis pipeline are comparable to the electrophoresis method. Additionally, the SGS method provides extra information and is able to call allele subtypes based on STR sequences in a database. In mixed samples, all alleles were reported from individuals that contributed at least 10 percent to the mixture. (Published Abstract)