Purification of forensic specimens for the polymerase chain reaction (PCR) analysis.
Purification methods of deoxyribonucleic acid (DNA) from degraded and contaminated forensic samples were investigated for polymerase chain reaction (PCR) analysis. DNA extracted from putrefied tissue or bloodstains sometimes contained the copurified contaminant, that was identified as the porphyrin compound (hematin). When contaminated but less degraded DNA was analyzed by PCR, it was necessary to eliminate the impurity by anion exchange column chromatography or chelating resin preparation, and ultrafiltration using Centricon microconcentrators. When highly degraded DNA was analyzed, trace amounts of high molecular weight DNA was recovered by electroelution method, and then further purified by both column chromatography and ultrafiltration. From thus purified samples, the amelogenin gene for sex determination could be amplified by dual PCR technique.
Akane
A
A
Department of Legal Medicine, Shimane Medical University, Izumo, Japan.
Shiono
H
H
Matsubara
K
K
Nakamura
H
H
Hasegawa
M
M
Kagawa
M
M
D000273:Adipose Tissue / Q000737:chemistry; D001801:Blood Stains; D004247:DNA / Q000032:analysis*; D005260:Female; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D008099:Liver / Q000737:chemistry; D008297:Male; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods*; D011180:Postmortem Changes
D004247:DNA