Use of a multiplex polymerase chain reaction assay in the sex typing of DNA extracted from archaeological bone

Abstract

We developed an original method that allows the simultaneous molecular amplification and analysis of human chromosome X‐ and Y‐specific sequences from cortical compact bone using the polymerase chain reaction (PCR). The strategy is based on sequential amplification using external and internal primers. The targeted loci are SRY for chromosome Y and DXZ4 for chromosome X. Internal and external primer pairs for both loci were selected in order to allow the simultaneous amplification of short sequences of distinct molecular weight in the same reaction. Ethidium bromide stained PCR products were analysed on 2 per cent agarose gels. The presence of two distinct amplification bands permitted the fast and unequivocal identification of male bones, whereas female bones were identified by the presence of a single band. The technique was validated by comparative analysis of modern human DNAs of known sex and of DNAs from medieval bone samples whose sex could be determined using conventional anthropological methods. We then analysed bone samples from 40 burials found under the Church of St Maria Aprutiensis in Teramo, Abruzzo. These burials were dated to AD 800–1200 by 14C accelerated mass spectrometry radiometric analysis. Molecular analysis allowed the unambiguous identification of sex in all the samples studied. These results indicate that molecular techniques represent useful and often critical additions to anthropological studies of ancient human remains. © 1997 by John Wiley & Sons, Ltd.

Sex determination of early medieval individuals through nested PCR using a new primer set in the SRY gene

“…However, PCR products were not obtained in any tested sample, regardless of modifications of PCR conditions originally designed by Palmirotta et al [8]. In male samples, aDNA extraction was repeated four times and samples were subjected to DXZ4/SRY nested PCR amplification to exclude accidental occurrence.…”

A Reliable and Reproducible Technique for DNA Fingerprinting in Biorepositories: A Pilot Study from BioBIM

“…Markers with an average heterozygosity ≥0.30 in different ethnic groups were also selected. The gender-typing loci DXZ4 and SRY were included in the study based on our previous results (12, 13) (Tab. I).…”

Optimization of subsampling, decontamination, and DNA extraction of difficult peat and silt permafrost samples

“…Likewise, many decontamination methods have been systematically tested for ancient DNA studies of skeletal remains. Some of these methods, such as scraping 31 and disk sampling 32 , have been used for permafrost decontamination as well. However, other methods used for aDNA studies of bone, including UV irradiation 33 , and treatment with household bleach 34 have not been tested on permafrost intended for microbial work.…”