HPLC and UV-Spectrophotometry Examination of Ancient DNA and PCR Inhibitors in Old Human Remains

Abstract

The study of ancient human remains can be approached today through polymerase chain reaction (PCR) analysis, but one of the main difficulties involves overcoming strong PCR inhibitions. We studied skeletal remains from six archaeological sites. The remains studied include a total of 27 individuals and span between 2500 and 400 years before the present. DNA was extracted from 35 samples and sex-specific amelogenin sequences were amplified to identify the sex of the samples. Only consistent results for which molecular and anthropometrical data agreed were considered. Notwithstanding the success of the sex identification, the samples were analysed using ultraviolet (UV) spectrophotometry and high performance liquid chromatography (HPLC) in order to establish a rapid protocol to predict the possible presence and state of ancient DNA. A combination of UV and HPLC peaks seems to indicate a strong degradation of endogenous DNA. A PCR inhibitor was also isolated through HPLC in a set of six samples.