Ancient Taurus cedar (Cedrus libani A. Rich) wood samples from the Tumulus of King Midas at the Gordion archaeological site (about 2700 years old), near Ankara, Turkey, and from the Al-Aksa Mosque (about 1500 to 1900 years old), Jerusalem, Israel, were characterized by studying the sequences of ribosomal DNA (rDNA) internal transcribed spacers (ITS1 and ITS2). After extraction of the DNA, the ITS regions were amplified utilizing the polymerase chain reaction, followed by sequencing, BLAST searches for similar sequences, and phylogenetic analyses. Fifty-six sequences were obtained.-In BLAST searches of existing sequence databases, most were closest to those from humans and fungi. However, two sequences exhibited similarities with conifer ITS sequences. One was an ITS1 region from the Gordion wood specimen, and the other one was an ITS2 region from the Al-Aksa wood specimen. Phylogenetic analyses indicated that both were closest to Taurus cedar (C. libani, also known as Lebanon cedar) ITS sequences from three recent samples of Taurus cedar from two sites in Turkey. However, they exhibited many differences from the recent C. libani rDNA ITS sequences from Turkey, probably due to degradation of the DNA in the ancient samples. The implications of the results on future studies are discussed.