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The Scientific World Journal
Volume 2013, Article ID 734676, 8 pages
http://dx.doi.org/10.1155/2013/734676
Research Article

Preamplification Procedure for the Analysis of Ancient DNA Samples

1Department of Experimental Medicine, Section of Biotechnology and Molecular Biology “A. Cascino”, Second University of Naples, Via Costantinopoli 16, 80138 Naples, Italy
2Department of Cardiothoracic Sciences, Second University of Naples, Via Leonardo Bianchi, 80131 Naples, Italy
3Anthropology Museum of Athens Medical School, Medical School, 75 M. Asias Street 11527 Goudi, Greece

Received 6 August 2013; Accepted 2 September 2013

Academic Editors: H. Kurumizaka and A. J. Piantino Ferreira

Copyright © 2013 Stefania Del Gaudio et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

In ancient DNA studies the low amount of endogenous DNA represents a limiting factor that often hampers the result achievement. In this study we extracted the DNA from nine human skeletal remains of different ages found in the Byzantine cemetery of Abdera Halkidiki and in the medieval cemetery of St. Spiridion in Rhodes (Greece). Real-time quantitative polymerase chain reaction (qPCR) was used to detect in the extracts the presence of PCR inhibitors and to estimate the DNA content. As mitochondrial DNA was detected in all samples, amplification of nuclear targets, as amelogenin and the polymorphism M470V of the transmembrane conductance regulator gene, yielded positive results in one case only. In an effort to improve amplification success, we applied, for the first time in ancient DNA, a preamplification strategy based on TaqMan PreAmp Master Mix. A comparison between results obtained from nonpreamplified and preamplified samples is reported. Our data, even if preliminary, show that the TaqMan PreAmp procedure may improve the sensitivity of qPCR analysis.