The Effects of Chemical and Heat Maceration Techniques on the Recovery of Nuclear and Mitochondrial DNA from Bone^*

Abstract

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.

Brief communication: Prenatal and early postnatal stress exposure influences long bone length in adult rat offspring

“…Bones were soaked at room temperature in a meat tenderizer solution for approximately 14 hours, manually cleaned of soft tissue, and allowed to dry. Soaking in meat tenderizer solution at up to 90°C is a common maceration procedure used to ensure complete removal of soft tissue for accurate bone measurement (Lee et al, 2010; Steadman et al, 2006). Maximum length of each bone was measured twice by the same person, who was blind to treatment group, to the nearest 0.01 mm using digital calipers.…”

Detection of the A189G mtDNA heteroplasmic mutation in relation to age in modern and ancient bones

“…First, the muscle was carefully removed from the bone with a scalpel. Then, the bones were placed in a microwave oven (5 x 1 min) to eliminate soft tissues [12]. The marrow remaining in the bone matrix was cleaned with distilled water and the bone was then crushed with a extremely cold, dry climate of this region generally provided a well-preserved nucleic acid chain [13].…”

A Simple Method to Extract DNA from Hair Shafts Using Enzymatic Laundry Powder

“…After extraction, extraction solutions were gradually heated up to 95°C to improve extract efficiency[10], and then subject to 95°C for 10 minutes in order to inactivate enzymes in the extraction reagent. The final DNA extracts were stored at −18°C until use.…”