title: Sequence variation of 22 autosomal STR loci detected by next generation sequencing.
abstract: Sequencing short tandem repeat (STR) loci allows for determination of repeat motif variations within the STR (or entire PCR amplicon) which cannot be ascertained by size-based PCR fragment analysis. Sanger sequencing has been used in research laboratories to further characterize STR loci, but is impractical for routine forensic use due to the laborious nature of the procedure in general and additional steps required to separate heterozygous alleles. Recent advances in library preparation methods enable high-throughput next generation sequencing (NGS) and technological improvements in sequencing chemistries now offer sufficient read lengths to encompass STR alleles. Herein, we present sequencing results from 183 DNA samples, including African American, Caucasian, and Hispanic individuals, at 22 autosomal forensic STR loci using an assay designed for NGS. The resulting dataset has been used to perform population genetic analyses of allelic diversity by length compared to sequence, and exemplifies which loci are likely to achieve the greatest gains in discrimination via sequencing. Within this data set, six loci demonstrate greater than double the number of alleles obtained by sequence compared to the number of alleles obtained by length: D12S391, D2S1338, D21S11, D8S1179, vWA, and D3S1358. As expected, repeat region sequences which had not previously been reported in forensic literature were identified.
authors.affiliation: U.S. National Institute of Standards and Technology, Biomolecular Measurement Division, 100 Bureau Drive, Gaithersburg, MD 20899, USA. Electronic address: katherine.gettings@nist.gov.
authors.forename: Katherine Butler
authors.initials: KB
authors.lastname: Gettings
authors.affiliation: U.S. National Institute of Standards and Technology, Biomolecular Measurement Division, 100 Bureau Drive, Gaithersburg, MD 20899, USA. Electronic address: kevin.kiesler@nist.gov.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Kiesler
authors.affiliation: North Carolina State University, College of Veterinary Medicine and Forensic Science Institute, 1060 William Moore Drive, Raleigh, NC 27607, USA. Electronic address: safaith@ncsu.edu.
authors.forename: Seth A
authors.initials: SA
authors.lastname: Faith
authors.affiliation: Battelle Memorial Institute, 505 King Avenue, Columbus, OH 43201, USA. Electronic address: montanoe@battelle.org.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Montano
authors.affiliation: Battelle Memorial Institute, 505 King Avenue, Columbus, OH 43201, USA. Electronic address: bakerc@battelle.org.
authors.forename: Christine H
authors.initials: CH
authors.lastname: Baker
authors.affiliation: Battelle Memorial Institute, 505 King Avenue, Columbus, OH 43201, USA. Electronic address: youngb@battelle.org.
authors.forename: Brian A
authors.initials: BA
authors.lastname: Young
authors.affiliation: Battelle Memorial Institute, 505 King Avenue, Columbus, OH 43201, USA. Electronic address: guerrierir@battelle.org.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Guerrieri
authors.affiliation: U.S. National Institute of Standards and Technology, Biomolecular Measurement Division, 100 Bureau Drive, Gaithersburg, MD 20899, USA. Electronic address: peter.vallone@nist.gov.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Vallone
mesh_terms: D001741:Black or African American / Q000235:genetics; D000483:Alleles; D004247:DNA / Q000032:analysis / Q000235:genetics; D053803:Forensic Genetics / Q000379:methods*; D006630:Hispanic or Latino / Q000235:genetics; D006801:Humans; D018895:Microsatellite Repeats; D044469:Racial Groups / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*; D044465:White People / Q000235:genetics
chemical_list: D004247:DNA
keywords: Next generation sequencing; Population genetics; STR