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Systematic Assessment of the Performance of Illumina's MiSeq FGx((TM)) Forensic Genomics System

NCJ Number
251946
Date Published
January 2017
Author(s)
Nouran Almalki, Hoi Yan Chow, Vishakha Sharma, Katie Hart, Donald Siegel, Elisha Wurmbach
Agencies
NIJ-Sponsored
Annotation
This study assessed the performance of Illumina's MiSeq FGx System for forensic genomics by systematically analyzing single source samples, evaluating concordance, sensitivity, and repeatability, as well as describing the quality of the reported outcomes.
Abstract
DNA from 16 individuals (9 males/7 females) in nine separate runs showed consistent STR profiles at DNA input =400 pg, and two full profiles were obtained with 50 pg DNA input; however, this study revealed that the outcome of a single sample does not merely depend on its DNA input but is also influenced by the total amount of DNA loaded onto the flow cell from all samples. Stutter and sequence or amplification errors can make the identification of true alleles difficult, particularly for heterozygous loci that show allele imbalance. Sequencing of 16 individuals’ STRs revealed genetic variations at 14 loci at frequencies suggesting improvement of mixture deconvolution. The STR loci D1S1656 and DXS10103 were most susceptible to drop-outs, and D22S1045 and DYS385a-b showed heterozygote imbalance. Most stutters were typed at TH01 and DYS385a-b, and amplification or sequencing errors were observed mostly at D7S820 and D19S433. Overall, Illumina's MiSeq FGx System produced reliable and repeatable results. aSTRs showed fewer drop outs than the Y- and X-STRs. (Publisher abstract modified)
Date Created: October 29, 2018