Purification of forensic specimens for the polymerase chain reaction (PCR) analysis
Akane, A.; Shiono, H.; Matsubara, K.; Nakamura, H.; Hasegawa, M.; Kagawa, M.
Journal of Forensic Sciences 38(3): 691-701
1993
ISSN/ISBN: 0022-1198 PMID: 8515220 Document Number: 409692
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.