Multiplex amplification of mitochondrial DNA for human and species identification in forensic evaluation

We describe a method combining in a single-round polymerase chain reaction amplifications of both cytochrome b and hypervariable D-loop mitochondrial DNA allowing species determination and individual human identification. Fol lowing the amplification step, amplicons are first screened on an agarose g el. The presence of only one band indicates that the sample is nonhuman, wh ile the presence of two bands indicates a human origin. Subsequent DNA sequencing of the hypervariable D-loop region DNA allows for individual human identification as the presence of cytochrome b fragment d oes not interfere with the analysis. Similarly, further species determinati on on the basis of the phylogenetically variable cytochrome b gene is possi ble by sequencing of the cytochrome b DNA fragment. (C) 1999 Elsevier Scien ce Ireland Ltd. All rights reserved.