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Document Type |
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Article In Journal |
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Document Title |
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Evaluation of the Illumina® beta version ForenSeq™ DNA signature prep kit for use in genetic profiling Evaluation of the Illumina® beta version ForenSeq™ DNA signature prep kit for use in genetic profiling |
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Document Language |
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English |
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Abstract |
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While capillary electrophoresis-based technologies have been the mainstay for human identity typing
applications, there are limitations with this methodology’s resolution, scalability, and throughput.
Massively parallel sequencing (MPS) offers the capability to multiplex multiple types of forensically-
relevant markers and multiple samples together in one run all at an overall lower cost per nucleotide than
traditional capillary electrophoresis-based methods; thus, addressing some of these limitations. MPS
also is poised to expand forensic typing capabilities by providing new strategies for mixture
deconvolution with the identification of intra-STR allele sequence variants and the potential to generate
new types of investigative leads with an increase in the overall number and types of genetic markers
being analyzed. The beta version of the Illumina ForenSeq DNA Signature Prep Kit is a MPS library
preparation method with a streamlined workflow that allows for targeted amplification and sequencing
of 63 STRs and 95 identity SNPs, with the option to include an additional 56 ancestry SNPs and 22
phenotypic SNPs depending on the primer mix chosen for amplification, on the MiSeq desktop sequencer
(Illumina). This study was divided into a series of experiments that evaluated reliability, sensitivity of
detection, mixture analysis, concordance, and the ability to analyze challenged samples. Genotype
accuracy, depth of coverage, and allele balance were used as informative metrics for the quality of the
data produced. The ForenSeq DNA Signature Prep Kit produced reliable, reproducible results and
obtained full profiles with DNA input amounts of 1 ng. Data were found to be concordant with current
capillary electrophoresis methods, and mixtures at a 1:19 ratio were resolved accurately. Data from the
challenged samples showed concordant results with current DNA typing methods with markers in
common and minimal allele drop out from the large number of markers typed on these samples. This set
of experiments indicates the beta version of the ForenSeq DNA Signature Prep Kit is a valid tool for
forensic DNA typing and warrants full validation studies of this MPS technology. |
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ISSN |
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1872-4973 |
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Journal Name |
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Forensic Science International: Genetics |
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Volume |
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20 |
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Issue Number |
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1 |
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Publishing Year |
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1437 AH
2016 AD |
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Article Type |
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Article |
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Added Date |
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Monday, July 10, 2017 |
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Researchers
| Jennifer D Churchill | Churchill, Jennifer D | Researcher | Doctorate | |
| Sarah E Schmedes | Schmedes, Sarah E | Researcher | Doctorate | |
| Jonathan L. King | King, Jonathan L. | Researcher | Doctorate | |
| Bruce Budowle | Budowle, Bruce | Researcher | Doctorate | |
|