Quantification and presence of human ancient DNA in burial place remains of Turkey using real time polymerase chain reaction

Abstract

Archaeometry and forensic laboratories are increasingly confronted with problematic samples from the scene of samples, containing only minute amounts of deoxyribonucleic acid (DNA), which may include
polymerase chain reaction (PCR) inhibiting substances. Efficient DNA extraction procedures, as well as accurate DNA quantification methods, are critical steps involved in the process of successful DNA analysis of such samples. Genomic DNA was extracted automatically by using EZ1 Automatic Nucleic Acid Isolation System (Qiagen, Germany) with investigator kit (Qiagen, Ilden, Germany) from ancient bones. This method is a sensitive for the extraction of DNA from a wide variety of forensic samples,
although it is known to be laborious compared with single tube extraction methods. The relatively high DNA recovery and the quality of the extracted DNA speak for itself. For reliable and sensitive DNA quantitation, the application of real time PCR is described. A published real-time PCR assay, which allows for the combined analysis of nuclear or ancient DNA and mitochondrial DNA, was modified. This approach can be used for recovering DNA from the surface of fossil bone remains in Turkey via a
simple procedure that permits a direct quantitative and qualitative assessment of molecular markers. Using quantitative RT-PCR, the available sources of total aDNA was shown to consists of intact DNA
that is virtually free of RNA, resulting in a more accurate representation of gene expression using RTPCR and PCR amplification methods. In this study, the results demonstrate that RT-PCR method can be
useful for the improved ancient DNA extraction in anthropology and archeology.