Preamplification Procedure for the Analysis of Ancient DNA Samples
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...
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| Format: | Article |
| Language: | English |
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Wiley
2013-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2013/734676 |
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| author | Stefania Del Gaudio Alessandra Cirillo Giovanni Di Bernardo Umberto Galderisi Theodoros Thanassoulas Theodoros Pitsios Marilena Cipollaro |
| author_facet | Stefania Del Gaudio Alessandra Cirillo Giovanni Di Bernardo Umberto Galderisi Theodoros Thanassoulas Theodoros Pitsios Marilena Cipollaro |
| author_sort | Stefania Del Gaudio |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-40336ecb471a47a09e727fa89e9e5839 |
| institution | Kabale University |
| issn | 1537-744X |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-40336ecb471a47a09e727fa89e9e58392025-02-03T07:25:43ZengWileyThe Scientific World Journal1537-744X2013-01-01201310.1155/2013/734676734676Preamplification Procedure for the Analysis of Ancient DNA SamplesStefania Del Gaudio0Alessandra Cirillo1Giovanni Di Bernardo2Umberto Galderisi3Theodoros Thanassoulas4Theodoros Pitsios5Marilena Cipollaro6Department of Experimental Medicine, Section of Biotechnology and Molecular Biology “A. Cascino”, Second University of Naples, Via Costantinopoli 16, 80138 Naples, ItalyDepartment of Experimental Medicine, Section of Biotechnology and Molecular Biology “A. Cascino”, Second University of Naples, Via Costantinopoli 16, 80138 Naples, ItalyDepartment of Experimental Medicine, Section of Biotechnology and Molecular Biology “A. Cascino”, Second University of Naples, Via Costantinopoli 16, 80138 Naples, ItalyDepartment of Experimental Medicine, Section of Biotechnology and Molecular Biology “A. Cascino”, Second University of Naples, Via Costantinopoli 16, 80138 Naples, ItalyDepartment of Cardiothoracic Sciences, Second University of Naples, Via Leonardo Bianchi, 80131 Naples, ItalyAnthropology Museum of Athens Medical School, Medical School, 75 M. Asias Street 11527 Goudi, GreeceDepartment of Experimental Medicine, Section of Biotechnology and Molecular Biology “A. Cascino”, Second University of Naples, Via Costantinopoli 16, 80138 Naples, ItalyIn 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.http://dx.doi.org/10.1155/2013/734676 |
| spellingShingle | Stefania Del Gaudio Alessandra Cirillo Giovanni Di Bernardo Umberto Galderisi Theodoros Thanassoulas Theodoros Pitsios Marilena Cipollaro Preamplification Procedure for the Analysis of Ancient DNA Samples The Scientific World Journal |
| title | Preamplification Procedure for the Analysis of Ancient DNA Samples |
| title_full | Preamplification Procedure for the Analysis of Ancient DNA Samples |
| title_fullStr | Preamplification Procedure for the Analysis of Ancient DNA Samples |
| title_full_unstemmed | Preamplification Procedure for the Analysis of Ancient DNA Samples |
| title_short | Preamplification Procedure for the Analysis of Ancient DNA Samples |
| title_sort | preamplification procedure for the analysis of ancient dna samples |
| url | http://dx.doi.org/10.1155/2013/734676 |
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