Ancient DNA extraction from bones and teeth.
This method is designed to maximize recovery of PCR-amplifiable DNA from ancient bone and teeth specimens and at the same time to minimize co-extraction of substances that inhibit PCR. This is achieved by a combination of DNA extraction from bone powder using a buffer consisting solely of EDTA and proteinase K, and purification of the DNA by binding to silica in the presence of high concentrations of guanidinium thiocyanate. All steps are performed at room temperature (20-23 degrees C), thereby reducing further degradation of the already damaged and fragile ancient DNA and providing an optimal trade-off between DNA release and degradation. Furthermore, the purification step removes most of the various types of PCR inhibitors present in ancient bone samples, thereby optimizing the amount of ancient DNA available for subsequent enzymatic manipulation, such as PCR amplification. The protocol presented here allows DNA extraction from ancient bone and teeth with a minimum of working steps and equipment and yields DNA extracts within 2 working days.
Rohland
Nadin
N
Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, D-04103 Leipzig, Germany. rohland@eva.mpg.de
Hofreiter
Michael
M
D000818:Animals; D001842:Bone and Bones / Q000737:chemistry*; D002498:Centrifugation / Q000379:methods; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D005580:Fossils; D005615:Freezing; D007202:Indicators and Reagents; D016133:Polymerase Chain Reaction / Q000379:methods; D012822:Silicon Dioxide / Q000302:isolation & purification; D014070:Tooth / Q000737:chemistry*
D007202:Indicators and Reagents; D012822:Silicon Dioxide; D004247:DNA