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DNA Extraction Method of Small Quantities of Bone for High-Quality Genotyping

NCJ Number
246419
Journal
Forensic Science International: Genetics Volume: 7 Issue: 5 Dated: September 2013 Pages: 488-493
Author(s)
Mariela Caputo; Maximiliano Irisarri; Evguenia Alechine; Daniel Corach
Date Published
September 2013
Length
6 pages
Annotation
DNA genotyping techniques have been used successfully in forensic science for almost three decades and represent the gold standard for individual identification.
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 1000 mg. We obtained optimal DNA yields 27 pg mg -1 on average when 150-200 mg 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.