This study assesses alternative methods of DNA recovery of compromised skeletal remains.
The results of this dissertation demonstrate significant and compelling differences between the treatment groups for the model moderately aged bovine and 19th-century human skeletal samples. The findings exemplify the issue of method-induced DNA degradation within samples processed by pulverization. Homogenizing skeletal elements into a fine powder dramatically impacts the recoverable template DNA, as shown in both length-based and targeted sequencing results from historical skeletal remains. Extended demineralization followed by slicing presents an alternative option for skeletal samples at the start of the forensic DNA analysis workflow. This study developed and optimized a prolonged demineralization and slicing method for small cortical bone fragments of bovine bone. Using demineralized slices resulted in more complete, balanced STR profiles, higher peak heights, and less degradation than powdered samples. In addition, a portion of this work evaluated chemical excision and physical cell capture methods using demineralized, collagenase-digested bovine cortical bone slices as a powder-free alternative for cell isolation. Ultimately, it was evident that the combined approach of collagenase treatment of demineralized slices did not improve the total yields of recovered DNA or STR profile development from aged and weathered bovine cortical bone samples. Similarly, employing micromanipulation methods on the partially digested demineralized bone slices did not increase endogenous DNA or reduce exogenous DNA yields in the samples. The efficacy of the developed demineralized slices method was assessed using human cortical skeletal samples in a comparative analysis with pulverized samples from the same donors. This study investigated the concordance of genetic markers through sequencing technologies to enhance the recovery of degraded DNA for human identification. These data showcase the effectiveness of a modified processing method for forensic identification through successful STR profile generation from skeletal remains aged over a century.