Restriction site mapping of the mitochondrial DNA of the four-wing flyingfish, Hirundichthys affinis

Mitochondrial DNA was isolated from samples of the four-wing flyingfish, Hi rundichthys affinis, collected in Barbados in January 1996 and subjected to restriction enzyme analysis, using 13 restriction endonucleases which reco gnise hexanucleotide sequences, in single and double digests. The resulting restricted DNA fragments were used to map the 14 enzyme recognition sites of 6 endonucleases (7 had no sites) on the flyingfish mtDNA molecule for th e first time. In addition, the mtDNA D-loop region was positioned on the re striction site map, for the first time, by selective restriction digestion of the mtDNA molecule followed by polymerase chain reaction (PCR) amplifica tion of the resulting fragments using specific mtDNA D-loop primers. The si ze of the flyingfish mtDNA molecule (18 kb) was also determined.