Contributions to Zoology, 77 (1) – 2008Ross Barnett; Nobuyuki Yamaguchi; Beth Shapiro; Richard Sabin: Ancient DNA analysis indicates the first English lions originated from North Africa
Materials and methods

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Data authenticity and analysis

Extraction of specimens NHM1952.10.20.15 and NHM 1952.10.20.16 took place in the ABC and was performed along with negative extraction controls. PCR amplification of a small hypervariable fragment of the mitochondrial control region was performed twice for each sample, each time incorporating negative amplification controls. The four resulting amplification products were then cloned using the TOPO TA system (Invitrogen, Carlsbad CA USA), sequenced on ABI377 automated sequencers (Perkin-Elmer, Wellesley MA USA), and aligned with previously published lion sequences (Barnett et al., 2006a, 2006b). A summary of the cloning results is presented in the Appendix. A total of 12 clones were sequenced from sample NHM 1952.10.20.15, and 10 from NHM1952. 10.20.16. Of these, only three sequences show evidence of postmortem DNA damage (E4, E6, and F5 in Table S1) and, in each instance, the damage occurs at nucleotide sites that are not known to be polymorphic in lions. A median-joining network was constructed from the resulting sequences using Network v4.1.0.3 (Bandelt et al., 1999).