The effects of chemical and heat maceration techniques on the recovery of nuclear and mitochondrial DNA from bone.
Forensic anthropologists use a number of maceration techniques to facilitate skeletal analysis of personal identity and trauma, but they may unwittingly eliminate valuable DNA evidence in the process. This study evaluated the effect of 10 maceration methods on gross bone structure and the preservation of DNA in ribs of 12 pigs (Sus scrofa). A scoring system was applied to evaluate the ease of maceration and resulting bone quality while DNA purity was quantified by optical densitometry analysis, followed by polymerase chain reaction (PCR) amplification of three mitochondrial and three nuclear loci. The results demonstrated that while mitochondrial DNA could be amplified for all experiments, cleaning treatments using bleach, hydrogen peroxide, ethylenediaminetetraacetic acid/papain, room temperature water and detergent/sodium carbonate followed by degreasing had low DNA concentrations and failed to generate nuclear PCR products. In general, treatments performed at high temperatures (90 degrees C or above) for short durations performed best. This study shows that traditionally "conservative" maceration techniques are not necessarily the best methods to yield DNA from skeletal tissue.
Steadman
Dawnie Wolfe
DW
Department of Anthropology, Binghamton University, SUNY, PO Box 6000, Binghamton, NY 13902-6000, USA. osteo@binghamton.edu
DiAntonio
Lisa L
LL
Wilson
Jeremy J
JJ
Sheridan
Kevin E
KE
Tammariello
Steven P
SP
D000818:Animals; D002254:Carbonates; D002614:Chelating Agents; D004247:DNA / Q000302:isolation & purification*; D017931:DNA Primers; D003902:Detergents; D004202:Disinfectants; D004492:Edetic Acid; D005503:Food Additives; D018732:Forensic Anthropology / Q000379:methods*; D006861:Hydrogen Peroxide; D007101:Immersion; D008872:Microwaves; D009812:Odorants; D016877:Oxidants; D010206:Papain; D016133:Polymerase Chain Reaction; D012272:Ribs / Q000473:pathology*; D017422:Sequence Analysis, DNA; D012973:Sodium Hypochlorite; D012997:Solvents; D013552:Swine; D013696:Temperature
D002254:Carbonates; D002614:Chelating Agents; D017931:DNA Primers; D003902:Detergents; D004202:Disinfectants; D005503:Food Additives; D016877:Oxidants; D012997:Solvents; C005686:sodium carbonate; D004247:DNA; D004492:Edetic Acid; D006861:Hydrogen Peroxide; D012973:Sodium Hypochlorite; D010206:Papain