A DNA extraction method of small quantities of bone for high-quality genotyping.
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 (27pg mg(-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.
Servicio de Huellas Digitales Genéticas, School of Pharmacy and Biochemistry, Universidad de Buenos Aires, Buenos Aires, Argentina. mcaputo@ffyb.uba.ar
Mariela
M
Caputo
Maximiliano
M
Irisarri
Evguenia
E
Alechine
Daniel
D
Corach
D053523:Amelogenin / Q000235:genetics; D001842:Bone and Bones / Q000737:chemistry*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D053803:Forensic Genetics / Q000379:methods*; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D018895:Microsatellite Repeats; D060885:Multiplex Polymerase Chain Reaction; D011208:Powders
D053523:Amelogenin; D011208:Powders; D004247:DNA
Exhumed-bone DNA extraction; Expert software; STR profile