Highlights
- •Multiplex STR typing of 17 STRs and the Amelogenin locus is feasible by MPS.
- •Sensitivity of detection is comparable with that of CE-based analyses
- •A size selection method and quantity of amplified DNA necessary are described.
- •The prototype STR multiplex and the illumina MiSeq generated reliable DNA profiles.
- •Intra-repeat SNPs were observed and this additional variation may be useful.
Abstract
STR typing in forensic genetics has been performed traditionally using capillary electrophoresis
(CE). However, CE-based method has some limitations: a small number of STR loci can
be used; stutter products, dye artifacts and low level alleles. Massively parallel
sequencing (MPS) has been considered a viable technology in recent years allowing
high-throughput coverage at a relatively affordable price. Some of the CE-based limitations
may be overcome with the application of MPS. In this study, a prototype multiplex
STR System (Promega) was amplified and prepared using the TruSeq DNA LT Sample Preparation
Kit (Illumina) in 24 samples. Results showed that the MinElute PCR Purification Kit
(Qiagen) was a better size selection method compared with recommended diluted bead
mixtures. The library input sensitivity study showed that a wide range of amplicon
product (6–200 ng) could be used for library preparation without apparent differences in the STR
profile. PCR sensitivity study indicated that 62 pg may be minimum input amount for generating complete profiles. Reliability study
results on 24 different individuals showed that high depth of coverage (DoC) and balanced
heterozygote allele coverage ratios (ACRs) could be obtained with 250 pg of input DNA, and 62 pg could generate complete or nearly complete profiles. These studies indicate that
this STR multiplex system and the Illumina MiSeq can generate reliable STR profiles
at a sensitivity level that competes with current widely used CE-based method.
Keywords
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Article info
Publication history
Published online: December 03, 2014
Accepted:
November 26,
2014
Received in revised form:
October 20,
2014
Received:
August 22,
2014
Identification
Copyright
© 2014 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
