Additional sequence characterization of NIST SRM 2391c: PCR-Based DNA Profiling Standard

Carolyn R. Hill, Margaret C. Kline, Michael Dewitt Coble, Peter M. Vallone

Research output: Contribution to journalArticle

Abstract

The NIST Standard Reference Material (SRM) 2391c: PCR-Based DNA Profiling Standard was designed for use in the standardization of forensic and paternity quality assurance procedures for fragment-based typing short tandem repeat (STR) alleles generated by the polymerase chain reaction (PCR). Certified genotypes of the 6 components A-F were assigned for 24 autosomal and 17 Y-STR markers plus Amelogenin using concordance testing between commercial kits. Selected Sanger sequencing characterization was performed for the alleles of 11 STR markers when only one PCR primer set was available for fragment-based typing. The goal is to characterize the remaining 30 STR loci in components A-C by Sanger sequencing methods for the STR repeat regions and adjacent flanking regions. Additional characterization of the SRM is intended to support the emerging interest in next-generation sequencing technologies for forensic typing applications. Sanger methods have detected underlying polymorphisms (sequence, insertion-deletion, variation in complex motifs) typically not detected by fragment-based typing. The sequenced regions include the commercial or known PCR binding sites commonly implemented in fragment-based typing.

Original languageEnglish
JournalForensic Science International: Genetics Supplement Series
Volume4
Issue number1
DOIs
StatePublished - 29 Oct 2013

Fingerprint

DNA Fingerprinting
Microsatellite Repeats
Polymerase Chain Reaction
Alleles
Amelogenin
Sequence Deletion
Insertional Mutagenesis
Binding Sites
Genotype
Technology

Keywords

  • DNA sequencing
  • DNA typing
  • Sanger sequencing
  • Short tandem repeat
  • Single nucleotide polymorphism
  • Standard reference material

Cite this

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abstract = "The NIST Standard Reference Material (SRM) 2391c: PCR-Based DNA Profiling Standard was designed for use in the standardization of forensic and paternity quality assurance procedures for fragment-based typing short tandem repeat (STR) alleles generated by the polymerase chain reaction (PCR). Certified genotypes of the 6 components A-F were assigned for 24 autosomal and 17 Y-STR markers plus Amelogenin using concordance testing between commercial kits. Selected Sanger sequencing characterization was performed for the alleles of 11 STR markers when only one PCR primer set was available for fragment-based typing. The goal is to characterize the remaining 30 STR loci in components A-C by Sanger sequencing methods for the STR repeat regions and adjacent flanking regions. Additional characterization of the SRM is intended to support the emerging interest in next-generation sequencing technologies for forensic typing applications. Sanger methods have detected underlying polymorphisms (sequence, insertion-deletion, variation in complex motifs) typically not detected by fragment-based typing. The sequenced regions include the commercial or known PCR binding sites commonly implemented in fragment-based typing.",
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Additional sequence characterization of NIST SRM 2391c : PCR-Based DNA Profiling Standard. / Hill, Carolyn R.; Kline, Margaret C.; Coble, Michael Dewitt; Vallone, Peter M.

In: Forensic Science International: Genetics Supplement Series, Vol. 4, No. 1, 29.10.2013.

Research output: Contribution to journalArticle

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