engleski

Application of the PowerPlex®S5 MINISTR System in Forensic DNA Analysis of Human Telogenic Hair Shafts

The primary value of DNA typing has been significantly increased over the last fifteen years due to introduction of short tandem repeat (STR) loci in paternity testing, as well as in forensic cases and mass disaster human identification. Nowadays, several commercial multiplex kits are available, allowing routine use of this powerful forensic-genetic tool. In addition, it is well known that even extremely highly degraded nuclear DNA may be successfully analyzed using PCR primers that are close to the STR repeat. This modification of STR molecular markers is described as miniSTR approach. In this study we have examined possible application of nuclear miniSTR loci in forensic DNA analysis of 65 telogenic hair that collected from various locations within the facility of Institute for Genetic Engineering and Biotechnology. All samples were briefly washed in absolute ethanol and extracted using Qiagen DNA Dnaeasy TM Tissue Kit. DNA concentration was determined using Quantifiler Human DNA Quantification Kit as described previously. The reaction was carried out in AB 7300 Real-Time PCR System according to the manufacturer’s recommendations. The PowerPlex S5® system has been used to amplify 4 miniSTR loci in 10µl total reaction volume. PCR amplification was carried out in AB GeneAmp PCR System Thermal Cycler. Electrophoresis of the amplification products was preformed on an ABI PRISM 310 Genetic Analyzer. Raw data have been compiled, analyzed and numerical allele designations of the profiles were obtained using the accessory software: ABI PRISM®, Data Collection Software v3.0 and GeneMapper TM ID Software 3.1. As the result of these analyses, 19 telogenic hair were completely or partially profiled and for 46 of them no profiles were detected. The most stable results were detected at TH01 locus. These facts, together with the other results of this study, imply that PowerPlex® S5 System may be useful in forensic analysis of the samples with very low amount of nuclear DNA such as telogenic hair. However, its application has certain limitations in the field that should also be considered and over- come in the future application of mini STR approach in the field of forensic genetic.

telogen hair; miniSTR; forensicDNA analysis

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