Abstract
BackgroundThe extraction of DNA from skeletal remains with good quality and quantity is often challenging for the ability to generate DNA typing. Previous studies demonstrated the DNA extraction with total demineralization from fresh teeth and bones; however, the application in old skeletal remains has been less performed. To obtain good quality and high yield of DNA amount extracted from skeletal remains, the objective of this study was focused on exploring the factors influencing the total demineralization process to obtain developing effective methods.ResultsThe concentration of EDTA was found to significantly enhance calcium chelation from the bone while pH of EDTA solution, incubation temperature, incubation time, and volume of EDTA solution were not significant. The optimal condition of total demineralization obtained from Placket-Burmann results represented good-quality DNA and the highest concentration of extracted DNA yield. Subsequently, the STR typing in some bone specimens processed by total demineralization process prior to DNA extraction was improved.ConclusionsEDTA concentration was a key influencing factor on the total demineralization process to chelate calcium from human skeletal remains. The total demineralization process in old bone specimens probably improved the STR profiles.
Highlights
The extraction of DNA from skeletal remains with good quality and quantity is often challenging for the ability to generate DNA typing
Total demineralization method based on Plackett-Burman design Due to the limited experimental samples when considering all factors into account, the program Design Expert version 6.0.8 was applied to find appropriate conditions
Each factor was determined at two levels, low (− 1) and high (+ 1) values, based on the literature reported on total demineralization of skeletal remains (Pérez et al 1989; Hagelberg and Clegg 1991; Żołędziewska et al 2002; Loreille et al 2007; Lee et al 2010; Bilic et al 2012)
Summary
The extraction of DNA from skeletal remains with good quality and quantity is often challenging for the ability to generate DNA typing. One of the crucial parts of the personal identification process from skeletal remains is based on the utility of the DNA profiles obtained from the bones or teeth (Latham and Miller 2019). Many techniques such as polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and gel electrophoresis have been extensively used to establish DNA profiling (Renneberg et al 2017). Seventy-five percent of the extracellular matrix was in the inorganic components consisting of calcium phosphate in the form of hydroxyapatite This substance supports the hard structure of the bone, resulting in limited DNA extraction (Loreille et al 2007). Several researchers have discovered the appropriate techniques to eliminate calcium phosphate from the bone structure (Pérez et al 1989; Serper and Çalt 2002; Sousa and Silva 2005; Sales et al 2018)
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