DNA and bone structure preservation in medieval human skeletons. (June 2015)

• Record Type: Journal Article
• Authors: Coulson-Thomas, Yvette M.; Norton, Andrew L.; Coulson-Thomas, Vivien J.; Florencio-Silva, Rinaldo; Ali, Nadir; Elmrghni, Samir; Gil, Cristiane D.; Sasso, Gisela R.S.; Dixon, Ronald A.; Nader, Helena B.
• Is Part Of: Forensic science international. Volume 251(2015)
• Page Start: 186
• Page End: 194
• Publication Date: 2015-06
• Subjects: Ancient DNA -- Archaeological bone -- Archaeological teeth -- Chelex -- DNA typing -- Electron microscopy; Medical jurisprudence -- Periodicals; Chemistry, Forensic -- Periodicals; Forensic Medicine -- Periodicals; Médecine légale -- Périodiques; Chimie légale -- Périodiques; Gerechtelijke geneeskunde; Gerechtelijke chemie; Gerechtelijke psychiatrie; Chemistry, Forensic; Medical jurisprudence; Electronic journals; Periodicals; Electronic journals; 614.1
• DOI: 10.1016/j.forsciint.2015.04.005
• Languages: English
• ISSNs: 0379-0738

Abstract

Highlights: Electron microscopy reveals the extracellular matrix is preserved in medieval bones. Medieval bone extracellular matrix proteins may be analyzed by immunohistochemistry. An efficient method is provided for isolating ancient DNA for forensic analysis. This adapted Chelex ® 100 method isolates high yields of archaeological skeleton DNA. Ancient DNA isolated using Chelex ® 100 is suitable for STR typing and real-time PCR. Abstract: Morphological and ultrastructural data from archaeological human bones are scarce, particularly data that have been correlated with information on the preservation of molecules such as DNA. Here we examine the bone structure of macroscopically well-preserved medieval human skeletons by transmission electron microscopy and immunohistochemistry, and the quantity and quality of DNA extracted from these skeletons. DNA technology has been increasingly used for analyzing physical evidence in archaeological forensics; however, the isolation of ancient DNA is difficult since it is highly degraded, extraction yields are low and the co-extraction of PCR inhibitors is a problem. We adapted and optimised a method that is frequently used for isolating DNA from modern samples, Chelex ® 100 (Bio-Rad) extraction, for isolating DNA from archaeological human bones and teeth. The isolated DNA was analysed by real-time PCR using primers targeting the sex determining region on the Y chromosome (SRY) and STR typing using the AmpFlSTR ® Identifiler PCR Amplification kit. Our results clearly show the preservation of bone matrix in medieval bones and the presence of intact osteocytes with well preserved encapsulated nuclei. In addition, we show how effective Chelex ® 100 is for isolating ancient DNA from archaeological bones and teeth. This optimised method is suitable for STR typing using kits aimed specifically at degraded and difficult DNA templates since amplicons of up to 250 bp were successfully amplified.