Typing of Deoxyribonucleic Acid (DNA) Extracted from Compact Bone from Human Remains

Abstract

The application of deoxyribonucleic acid (DNA) typing methods for the potential identification of unknown human remains was investigated. DNA was isolated from compact bone tissue from badly decomposed bodies and from known and unknown human remains, using a decalcification and ion wash procedure. Restriction fragment length polymorphism (RFLP) analysis of variable number of tandem repeats (VNTR) loci yielded results in some cases, but more often the DNA was too degraded to produce RFLP patterns. No RFLP profiles could be obtained from putrified soft tissues. However, DNA extracted from compact bone tissue of human remains up to eleven years old was successfully amplified using the polymerase chain reaction (PCR) for the VNTR loci D1S80, D17S5, COL2A1, and APO B, as well as the HLA-DQ alpha locus. This is especially significant, since PCR results were obtained from those samples whose DNA had been degraded substantially and had yielded no RFLP patterns. All DNA types determined from the compact bone tissue from decomposed bodies whose identification had been established first by other means (and whose parents or offspring were available for typing) demonstrated mendelian inheritance of the alleles of the loci analyzed. These results suggest that amplification and typing of DNA extracted from compact bone of human remains could be useful in establishing the identity of a person, as well as in excluding possible false identifications.

Synergy between DNA polymerases increases polymerase chain reaction inhibitor tolerance in forensic DNA analysis

“…The presence or absence of PCR facilitators is a well-known factor determining the amplification efficiency of DNA polymerases in general [35], and several different compounds have been used to counteract PCR inhibition [25]. Examples of PCR facilitators that have been shown to increase inhibitor tolerance include proteins such as BSA [14,26] and nonionic detergents such as Tween 20 and Triton X-100 [33,36]. BSA is included in the AmpFlSTR reaction buffer used routinely with the AmpFlSTR SGM Plus kit (AmpFlSTR SGM Plus PCR Amplification Kit user's manual).…”

“…For most samples, DNA extraction was performed using Chelex beads (Bio-Rad Laboratories, Hercules, CA, USA) [12], with the addition of Centricon purification (Millipore, Billerica, MA, USA) [13] for visibly dirty samples. Phenolchloroform purification [14] was used on especially complicated samples. A complete list of which DNA extraction/purification method was used for each sample is provided in Table S2 of the Supplementary material.…”

An efficient DNA extraction method from bone and tooth samples by complete demineralization followed by the use of silica-based columns

“…In a mass disaster, recovery of STR data from bones and tooth exposed to environmental conditions has become a valuable tool for the identification of missing individuals (16) . In case of extreme degradation bone and tooth may be the only suitable material available for successful DNA typing.…”

Short Tandem Repeat Typing Using Endogenous DNA Isolated from Human Fingernail

“…One force characteristic of DNA profiling is that it may be used in assortment of various biological sources. Some of these sources include blood [1,2], semen [3], hair [4][5][6], saliva [7],bone [8][9][10][11][12][13][14] and teeth [15][16][17]. DNA profiling of individuals with short tandem repeat (STR) markers is commonly used in forensic genetics and paternity testing.…”