A DNA extraction method of small quantities of bone for high-quality genotyping

Abstract

DNA genotyping techniques have been used successfully in forensic science for almost three decades and represent the gold standard for individual identification. However, efficient protocols for obtaining DNA from exhumed bones suitable for genotyping are still scarce and most of them require a considerable amount of starting material, are time consuming and are inefficient for reducing inhibitor's effects. We sought to develop an optimised protocol for extracting DNA from bone samples obtained from exhumations. We tested two approaches for preparing bone samples: (a) fine powder and (b) thin slices of bone. The best ratio of bone amount to DNA yields was assessed by a titration experiment using bone powder ranging from 50 to 1000mg. We obtained optimal DNA yields (27pgmg(-1) on average) when 150-200mg of starting material were processed using a one-step demineralisation method. Better-quality profiles (determined by the number of genotyped loci) were obtained when DNA was extracted from bone slices compared to extraction from bone powder. From bone slices 83.9% and from bone powder 46.7% of the samples provided genotypes for 11 or more loci. Since bone preparation procedures were carried out at room temperature, the method developed in the present study might be an attractive alternative to the standard freeze-mill approach, being faster and more cost-efficient.