MBE Advance Access published online on July 14, 2004
Molecular Biology and Evolution, doi:10.1093/molbev/msh208
Molecular Biology and Evolution © Society for Molecular Biology and Evolution 2004; all rights reserved
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1 Institute of Anthropology and Human Genetics, Division of Molecular Genetics, University of Tübingen, Wilhelmstr. 27, D-72074 Tübingen, Germany
* To whom correspondence should be addressed. E-mail: bachmann{at}nhm.uio.no.
Achondroplasia (ACH) is a skeletal disorder (MIM100800) with an autosomal dominant Mendelian inheritance and complete penetrance. We here report the screening of ancient bone samples for diagnostic ACH mutations. The diagnostic G
Original Articles
PCR-Induced Sequence Alterations Hamper the Typing of Prehistoric Bone Samples for Diagnostic Achondroplasia Mutations
2 Institute of Anthropology and Human Genetics, Division of Molecular Genetics, University of Tübingen, Wilhelmstr. 27, D-72074 Tübingen, Germany; Institut für Ur- und Frühgeschichte, Abteilung Ältere Urgeschichte und Quartärökologie, University of Tübingen, Burgsteige 11, D-72070 Tübingen, Germany
3 Institute of Organic Chemistry, University of Tübingen, Auf der Morgenstelle 18, D-72076 Tübingen, Germany
4 Institute of Pathology, Division of Palaeopathology, Academic Teaching Hospital München-Bogenhausen, Englschalkingerstr. 77, D-81925 München, Germany
5 Institute of Human Genetics, University of Münster, Vesaliusweg 12-14, D-48149 Münster, Germany
6 Department of Zoology, Natural History Museums and Botanical Garden, University of Oslo, P.O.Box 1172 Blindern, N-0318 Oslo, Norway
Abstract 
A transition in the FGFR3 gene at cDNA position 1,138 was detected in cloned PCR products obtained from the dry mummy of the Semerchet tomb, Egypt, (first dynasty, 
4,890-5,050 BP) and an individual from Kirchheim, Germany, (Merovingian period, 1,300-1,500 BP) that both had short stature. However, these mutations were also reproducibly observed in four ancient control samples from phenotypically healthy individuals (false-positives) rendering the reliable molecular typing of ancient bones for ACH impossible. The treatment of a false-positive DNA extract with uracil N-glycosylase (UNG) in order to minimize type 2 transitions (GA/CT) did not reduce the frequency of the false-positive diagnostic ACH mutations. Recently, it was suggested that ancient DNA extracts may induce mutations under PCR. Contemporary human template DNA from a phenotypically healthy individual was therefore spiked with an ancient DNA extract from a cave bear. Again, sequences with the diagnostic GA transition in the FGFR3 gene were observed, and it is likely that the false-positive GA transitions result from errors introduced during the PCR reaction. Amplifications under the presence of MnCl2 indicate that position 1,138 of the FGFR3 gene is particularly sensitive for mutations. Our data are in line with previously published results on the occurrence of non-random mutations in PCR products of contemporary human mitochondrial HVRI template DNA spiked with ancient DNA extracts.
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