Developmental validation of a real-time PCR assay for the simultaneous quantification of total human and male DNA.
Multiplex human short tandem repeat analysis demands reliable DNA quantification to consistently produce interpretable genotypes. The Plexor HY System is a multiplex quantitative PCR assay to quantify total human and male DNA. We performed developmental validation of the Plexor HY System to demonstrate the performance capabilities and limitations of the assay for forensic applications. Validation studies examined: (a) human specificity, (b) sensitivity, (c) quantification of degraded DNA, (d) impact of inhibitors, (e) male/female mixture and Y-assay male specificity, (f) reproducibility and concordance and (g) population studies.
Krenke
Benjamin E
BE
Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, USA. benjamin.krenke@promega.com
Nassif
Nadine
N
Sprecher
Cynthia J
CJ
Knox
Curtis
C
Schwandt
Melissa
M
Storts
Douglas R
DR
D000818:Animals; D041322:Chromosomes, Human, Y / Q000235:genetics; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004271:DNA, Fungal / Q000235:genetics; D053803:Forensic Genetics / Q000379:methods; D005838:Genotype; D006801:Humans; D008297:Male; D008322:Mammals / Q000235:genetics; D018895:Microsatellite Repeats / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012727:Sex Characteristics; D014998:Y Chromosome / Q000235:genetics
D004271:DNA, Fungal; D004247:DNA