DNA and bone structure preservation in medieval human skeletons.
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 250bp were successfully amplified.
Department of Biochemistry, Universidade Federal de São Paulo, Rua Três de Maio 100, São Paulo, 04044-020, Brazil; School of Life Sciences, University of Lincoln, Brayford Pool, Lincoln, LN6 7TS, UK. Electronic address: ycoulsonthomas@gmail.com.
Yvette M
YM
Coulson-Thomas
Durham University, Woodland Road, DH7 9RH, Durham, UK.
Andrew L
AL
Norton
Department of Biochemistry, Universidade Federal de São Paulo, Rua Três de Maio 100, São Paulo, 04044-020, Brazil; John van Geest Centre for Brain Repair, University of Cambridge, Forvie Site, Robinson Way, Cambridge, CB2 0PY, UK.
Vivien J
VJ
Coulson-Thomas
Department of Morphology and Genetics, Universidade Federal de São Paulo, Rua Botucatu 740, São Paulo, 04023-900, Brazil.
Rinaldo
R
Florencio-Silva
School of Life Sciences, University of Lincoln, Brayford Pool, Lincoln, LN6 7TS, UK.
Nadir
N
Ali
School of Life Sciences, University of Lincoln, Brayford Pool, Lincoln, LN6 7TS, UK.
Samir
S
Elmrghni
Department of Morphology and Genetics, Universidade Federal de São Paulo, Rua Botucatu 740, São Paulo, 04023-900, Brazil.
Cristiane D
CD
Gil
Department of Morphology and Genetics, Universidade Federal de São Paulo, Rua Botucatu 740, São Paulo, 04023-900, Brazil.
Gisela R S
GR
Sasso
School of Life Sciences, University of Lincoln, Brayford Pool, Lincoln, LN6 7TS, UK.
Ronald A
RA
Dixon
Department of Biochemistry, Universidade Federal de São Paulo, Rua Três de Maio 100, São Paulo, 04044-020, Brazil.
Helena B
HB
Nader
D000328:Adult; D001842:Bone and Bones / Q000737:chemistry* / Q000648:ultrastructure*; D041322:Chromosomes, Human, Y; D024061:Collagen Type III / Q000032:analysis; D004247:DNA / Q000302:isolation & purification*; D016172:DNA Fingerprinting; D017931:DNA Primers; D005260:Female; D049691:History, Medieval; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D018895:Microsatellite Repeats; D046529:Microscopy, Electron, Transmission; D015675:Osteocalcin / Q000032:analysis; D010011:Osteocytes / Q000166:cytology; D060888:Real-Time Polymerase Chain Reaction; D012732:Sex Determination Analysis; D014070:Tooth / Q000737:chemistry
D024061:Collagen Type III; D017931:DNA Primers; D015675:Osteocalcin; D004247:DNA
Ancient DNA; Archaeological bone; Archaeological teeth; Chelex; DNA typing; Electron microscopy