FORENSIC SCIENCE INTERNATIONAL: GENETICS SUPPLEMENT SERIES
Abstract
A set of 16 amplifications following a D-optimal experimental design was planned using the most
commonly selected kit and sequencer utilized in a collaborative exercise of the ISFG Italian Working
Group GeFI on the molecular characterization of a depurinated DNA sample. The data were evaluated
using STRvalidator in order to describe the occurrence of PCR artifacts such as peak imbalance and allele/
locus drop outs. The aim of the study was to describe the role of the template amount, number of
amplification cycles, volume of the PCR reaction in determining the assay sensitivity and the profiling
accuracy as measured by peak heights and areas. The results showed that only by selecting an appropriate
ratio of template to PCR reaction solution total volume together with an increase in the number of cycles
it could be possible to obtain balanced peaks as well as minimal occurrence of other artifacts. These data
are useful to model and understand the PCR artifacts occurrence described in the collaborative exercise.
