TY - JOUR
T1 - Mitochondrial DNA Analysis in a 5300-year-old Specimen from Israel
AU - Agamy, L.
AU - Faerman, Marina
AU - Smith, P.
N1 - doi: 10.1080/1358612021000040430
PY - 2002
Y1 - 2002
N2 - We attempted DNA extraction from 14 bone and dental specimens found at Wadi Maqoch site, which is located in the Jordan Valley and dated to the Chalcolithic period. DNA was isolated from tiny amounts of bone and tooth powder following a slightly modified silica extraction protocol. Stringent precautions were undertaken to prevent possible contamination of the samples. Each DNA sample was subjected to a number of PCR reactions. Sex identification was performed using primers and conditions described by Sullivan and colleagues, but failed to provide positive results. Hypervariable segment I (HS-I) of the mitochondrial DNA control region was amplified in three overlapping PCR reactions. One of the DNA samples yielded amplification products of 153 bp, 232 bp and 271 bp that together comprised almost the complete HS-I. Automated sequencing revealed the following changes as compared to the Cambridge Reference Sequence: G to C at position 16,129, C to T at position 16,223. In addition, equal peaks of C and T were observed at position 16,311, suggesting possible heteroplasmy. Notably, the presence of heteroplasmy in position 16,311 has been already revealed in a number of studies. Cloning of PCR products will allow us to verify this finding. It is important to mention that two PCR reactions overlap at 16,223. We repeatedly observed a T at this position. Furthermore, none of the investigators in the laboratory carries this mtDNA haplotype or any of these polymorphisms. An extensive search in different mtDNA databases has revealed no exact matches to the sequence obtained in the Chalcolithic specimen. Three individuals (Moroccan Jew, Cantonese and Swiss) were found to differ at position 16,129 by possessing A and not C.
AB - We attempted DNA extraction from 14 bone and dental specimens found at Wadi Maqoch site, which is located in the Jordan Valley and dated to the Chalcolithic period. DNA was isolated from tiny amounts of bone and tooth powder following a slightly modified silica extraction protocol. Stringent precautions were undertaken to prevent possible contamination of the samples. Each DNA sample was subjected to a number of PCR reactions. Sex identification was performed using primers and conditions described by Sullivan and colleagues, but failed to provide positive results. Hypervariable segment I (HS-I) of the mitochondrial DNA control region was amplified in three overlapping PCR reactions. One of the DNA samples yielded amplification products of 153 bp, 232 bp and 271 bp that together comprised almost the complete HS-I. Automated sequencing revealed the following changes as compared to the Cambridge Reference Sequence: G to C at position 16,129, C to T at position 16,223. In addition, equal peaks of C and T were observed at position 16,311, suggesting possible heteroplasmy. Notably, the presence of heteroplasmy in position 16,311 has been already revealed in a number of studies. Cloning of PCR products will allow us to verify this finding. It is important to mention that two PCR reactions overlap at 16,223. We repeatedly observed a T at this position. Furthermore, none of the investigators in the laboratory carries this mtDNA haplotype or any of these polymorphisms. An extensive search in different mtDNA databases has revealed no exact matches to the sequence obtained in the Chalcolithic specimen. Three individuals (Moroccan Jew, Cantonese and Swiss) were found to differ at position 16,129 by possessing A and not C.
U2 - 10.1080/1358612021000040430
DO - 10.1080/1358612021000040430
M3 - Article
SN - 1358-6122
VL - 4
SP - 121
EP - 167
JO - Ancient Biomolecules
JF - Ancient Biomolecules
IS - 3
ER -