Extraction and amplification of nuclear and mitochondrial DNA from ancient and artificially aged bones.

ABSTRACT

An experimental design is presented that simulates an accelerated process of DNA degradation in human bone tissues and thus provides a possibility for a systematic investigation of factors hampering DNA extraction and amplification. Equal sized slices of human femoral bones were incubated in 90 degrees C water for 2 h up to 30 days. DNA was extracted and subjected to a human specific Duplex polymerase chain reaction (PCR) and also to a Multiplex short tandem repeat (STR) PCR. Additionally 24 ancient bones representing different age periods were investigated in the same way. The results were compared to those from the artificially aged samples. After just 12 h of incubation, DNA is totally degraded, but still fully typable. After 36 h no reproducible amplification of DNA is possible. Using Multiplex PCR the DNA from artificially aged bones shows the typical STR pattern for ancient samples suggesting that the in vitro approach provides a useful and comparable method to elucidate the DNA degradation process in bones.