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. Following the amplification step, amplicons are first screened on an agarose gel. The presence of only one band indicates that the sample is nonhuman, while 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 does not interfere with the analysis. Similarly, further species determination on the basis of the phylogenetically variable cytochrome b gene is possible by sequencing of the cytochrome b DNA fragment.
Bataille
M
M
Laboratoire de Biochimie et Biologie Moléculaire, Hôpital R. Poincaré, Garches, France.
Crainic
K
K
Leterreux
M
M
Durigon
M
M
de Mazancourt
P
P
D000818:Animals; D003573:Cytochrome b Group / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods*; D004272:DNA, Mitochondrial / Q000235:genetics*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013045:Species Specificity; D013552:Swine / Q000235:genetics
D003573:Cytochrome b Group; D004272:DNA, Mitochondrial; D007135:Immunoglobulin Variable Region