NCJ Number
226994
Journal
Forensic Science International: Genetics Volume: 3 Issue: 2 Dated: March 2009 Pages: 96-103
Date Published
March 2009
Length
8 pages
Annotation
This report presents the design of a new multiplex polymerase chain reaction (PCR) with reduced size amplicons (<200 bp), providing a double sex determination (amelogenin plus two Y-STRs), the detection of two autonomic markers, and the amplification of mitochondrial specific fragments from the hypervariable region I (HVI); a quality sensor was developed to check for the presence of any PCR inhibitors.
Abstract
This new multiplex PCR produces a reproducible detection threshold down to 25 pg and gives signals even out of highly degraded materials. All signals are reproducible and reliable. In no case were DNA fragments detectable using any other assay when the quality sensor was not detectable. There was a good correlation between detection of mitochondrial specific fragments in the multiplex PCR and the success of subsequent sequencing of HVI region. The same could be shown for short tandem repeat (STR) analysis. Most samples successfully analyzed in the PCR yielded at least a partial STR profile using a commercial STR kit. This new assay allows an easy, reliable, and cost-efficient evaluation of DNA sample quality, combined with a first rough sample individualization and sex determination suitable for forensic purposes. This assay can help the forensic lab personnel decide whether to pursue further sample processing. For PCR positive and contamination controls and for dilution experiments, a commercially available DNA was used (Human Genomic DNA female and male DNA, Promega, Germany). For quality control, DNA from 100 males and 50 females was extracted from saliva swabs or blood samples using the Spin Swab kit or the Nucleo Spin Blood Quick Pure kit, respectively. In addition, 127 samples from various biological materials that were part of the researchers' routine case analyses were investigated. 4 Tables, 4 figures and 44 references