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The study of DNA sequence data obtained from specimens of extinct organisms offers a window into the evolutionary past. However, this area of research is technically very challenging and is strewn with potential pitfalls. Handt et al. (1994) and others (Lindahl 1993; Paabo, Higuchi, and Wilson 1989) have emphasised the need for careful experimental design and procedure in ancient DNA work as well as the application of criteria of authenticity. Moreover, independent reproducibility of presumed ancient DNA sequence is essential if authenticity is to be properly evaluated. The reproducibility of such sequences is tested here by independent investigation of archaeological samples of Pleistocene fauna. A total of 13 samples of soft tissue and bone from three species of Pleistocene mammals were analysed. Two of these species, Mylodon darwinii and Mammuthus primigenius, are extinct. The status of the other specimen, a putative Equus sp., is unclear. Details of the samples are listed in table 1. Only one of these three species, Mammuthus primigenius, has previously been shown to yield DNA sequence in two independent studies (Hagelberg et al. 1994; H&s, Paabo, and Vereschagin 1994), albeit for different segments of the mitochondrial genome. No endogenous ancient DNA sequences have so far been published for either the Mylodon or the Selerikan horse. The region of the mitochondrial 16s ribosomal RNA gene previously studied by H&s, Paabo, and Vereschagin (1994) was selected as the target for PCR amplification in this study. All manipulations prior to thermal cycling (prePCR) were carried out in a laboratory physically separated and remote from procedures performed after PCR (post-PCR). Dedicated protective clothing, reagents, pipettes, gamma irradiated filter tips, and sterile disposable plasticware were used at all times during pre-PCR operations. Risk of contamination was further minimised by frequent use of 10% sodium hypochlorite solution to clean surfaces (Prince and Andrus 1992) and UV irradiation of tubes and solutions (Fox et al. 199 1; Ou,

Reproducibility of ancient DNA sequences from extinct Pleistocene fauna