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 language | English |
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Pages (from-to) | 220-227 |
Number of pages | 8 |
Journal | Forensic Science International |
Volume | 133 |
Issue number | 3 |
DOIs | |
State | Published - 5 May 2003 |
Keywords
- AmpFℓSTR ®
- D8S1179
- Multiplex
- PCR
- Primer binding site mutation
- STR
- Validation