A simple and efficient method for PCR amplifiable DNA extraction from ancient bones.
A simple and effective modified ethanol precipitation-based protocol is described for the preparation of DNA from ancient human bones. This method is fast and requires neither hazardous chemicals nor special devices. After the powdering and incubating of the bone samples Dextran Blue was added as a carrier for removing the PCR inhibitors with selective ethanol precipitation. This method could eliminate the time-consuming separate decalcification step, dialysis, application of centrifugation-driven microconcentrators and the second consecutive PCR amplification. The efficiency of this procedure was demonstrated on ten 500-1200-year-old human bones from four different Hungarian burial sites. A mitochondrial specific primer pair was used to obtain sequence information from the purified ancient DNA. The PCR amplification, after our DNA extraction protocol, was successful from each of the 10 bone samples investigated. The results demonstrate that extraction of DNA from ancient bone samples with this new approach increases the success rate of PCR amplification.
Kalmár
T
T
Institute of Genetics, Biological Research Centre of Hungarian Academy of Sciences, POB 521, H-6701, Szeged, Hungary and Department of Anthropology, University of Szeged, Egyetem u. 2, H-6725, Szeged, Hungary. klampar@nucleus.szbk.u-szeged.hu
Bachrati
C Z
CZ
Marcsik
A
A
Raskó
I
I
D000293:Adolescent; D000328:Adult; D000368:Aged; D001842:Bone and Bones / Q000737:chemistry*; D004247:DNA / Q000266:history / Q000302:isolation & purification*; D005260:Female; D049668:History, 15th Century; D049691:History, Medieval; D006801:Humans; D006814:Hungary; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*
D004247:DNA