Identification of a D8S1179 primer binding site mutation and the validation of a primer designed to recover null alleles

Craig Leibelt, Bruce Budowle, Patrick Collins, Yasser Daoudi, Tamyra Moretti, Gary Nunn, Dennis Reeder, Rhonda Roby

Research output: Contribution to journalArticle

66 Scopus citations

Abstract

A population study of Chamorros and Filipinos using short tandem repeat (STR) loci amplified with the AmpFℓSTR® Profiler Plus™ PCR amplification kit demonstrated an excess of observed homozygosity at the D8S1179 locus. Use of a different set of D8S1179 primers to type the same samples did not demonstrate an excess of homozygosity and showed discordant genotypes at the D8S1179 locus. A single point mutation, G-to-A transition, 16 nucleotides from the 3′ end of the reverse primer, was identified to cause allele dropout when using the AmpFℓSTR® Profiler Plus™ primer set. An additional D8S1179 reverse primer specific for the variant was constructed resulting in the recovery of the null allele. The primer was included in the newly developed AmpFℓSTR® Identifiler™ PCR amplification kit. No deleterious effects or non-specific peaks were observed in validation experiments evaluating primer concentration, Mg2+ concentration, annealing temperature and population samples.

Original languageEnglish
Pages (from-to)220-227
Number of pages8
JournalForensic Science International
Volume133
Issue number3
DOIs
StatePublished - 5 May 2003

Keywords

  • AmpFℓSTR ®
  • D8S1179
  • Multiplex
  • PCR
  • Primer binding site mutation
  • STR
  • Validation

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