Document Details

Document Type : Article In Journal 
Document Title :
High sensitivity multiplex short tandem repeat loci analyses with massively parallel sequencing
High sensitivity multiplex short tandem repeat loci analyses with massively parallel sequencing
 
Document Language : English 
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 (6-200ng) could be used for library preparation without apparent differences in the STR profile. PCR sensitivity study indicated that 62pg 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 250pg of input DNA, and 62pg 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. 
ISSN : 1878-0326 
Journal Name : Forensic Sci Int Genet 
Volume : 16 
Issue Number : 1 
Publishing Year : 1436 AH
2015 AD 
Article Type : Article 
Added Date : Sunday, May 1, 2016 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
Xiangpei ZengZeng, Xiangpei Investigator  
Jonathan L. KingKing, Jonathan L.Researcher  
Monika StoljarovaStoljarova, Monika Researcher  
David H. WarshauerWarshauer, David H.Researcher  
Bobby L. LaRueLaRue, Bobby L.Researcher  
Antti SajantilaSajantila, Antti Researcher  
Jaynish PatelPatel, Jaynish Researcher  
Douglas R. StortsStorts, Douglas R.Researcher  
Bruce BudowleBudowle, Bruce Researcher Bruce.Budowle@unthsc.edu

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