**3. Results**

The bone powder obtained during bone pre-processing was used for DNA extraction. In this case, bone demineralization was performed. The DNA concentrations of the saliva, petrous pyramid and tooth after quantification using the Power Quant® System (Promega, USA) are presented in **Tables 1**–**3** for STR markers.

Using capillary electrophoresis, we obtained the genotypes on STR markers as follows: in **Case 1 –** the paternity relationship between the presumptive son and his deceased father was confirmed (**Table 4**); **Case 2 –** the maternity relationship between the presumptive daughter and the victim found in the woods was proven (**Table 5**); **Case 3 –** the kinship between the unrecognizable burn victim and his presumptive daughter was established (**Table 6**).


#### **Table 4.**

*Genetic DNA profiles in Case 1. The paternal alleles are shown colored.*


#### *Genetic DNA Identification from Bone Remains in Kinship Analysis Using Automate… DOI: http://dx.doi.org/10.5772/intechopen.99587*

#### **Table 5.**

*Genetic DNA profiles in Case 2. The maternal alleles are shown colored.*


#### *Criminology and Post-Mortem Studies - Analyzing Criminal Behaviour and Making Medical…*


**Table 6.**

*Genetic DNA profiles in Case 3. The paternal alleles are shown colored.*

The Gene Mapper ID-X software version 1.4 (Thermo Fischer Scientific, USA) was used to analyze the data.

#### **4. Discussion**

Genetic analysis is a fundamental tool that is used for the identification of skeletonized remains. In forensic genetics, the most important steps for genetic identification are DNA extraction from biological samples and their quantification.

In our cases, from the biological samples that were analyzed: the saliva provided by the first-degree relatives of the deceased as reference sample in all cases, and a canine tooth, a femur bone fragment, and a petrous pyramid extracted from the victims, all had good concentrations on autosomal STR markers.

The DNA concentrations of the biological samples are presented in **Tables 1**–**3**. An important aspect to look out for while performing genetic identification of skeletonized remains relates to the relationship between the types of bones and the DNA concentration obtained during the analysis. Many studies have demonstrated that compact bones yield a greater amount of DNA when compared to spongy bones [11–13]. In **Tables 4**–**6**, the genetic profiles obtained from bones remains are presented together with the genetic profiles of the first degree relatives, thus being demonstrated the kinship relationship between them. In these studies, it was demonstrated that compact bones from the lower limbs are more effective as DNA concentrations when compared to compact bones from the upper limbs [14].

Many studies classified the types of human bones that are recommended for forensic genetic identification depending on the amount of DNA that can be obtained, such as: tooth, talus, tarsal bones, petrous temporal bone, vertebra, femur and tibial metatarsal [15]. In our cases, our results conformed to the results obtained by other forensic genetics laboratories, in that the DNA concentrations differed

#### *Genetic DNA Identification from Bone Remains in Kinship Analysis Using Automate… DOI: http://dx.doi.org/10.5772/intechopen.99587*

based on the bone type. The tooth was observed to have the greatest DNA concentration when compared to the petrous temporal bone or the femur bone.

During bone demineralization, some laboratories had included liquid nitrogen in their grinding procedure and mill freeze and bone powder incubation was observed for 72 hours [16–18]. Following the advances made in molecular techniques for DNA extraction from skeletonized remains, in 2019, Promega Company introduced a new kit for DNA extraction from highly degraded bones. This kit optimized the process of DNA extraction and PCR amplification, thereby enabling forensic laboratories to obtain good results while causing minimal destruction to bone samples and using a minimal time of only three hours for DNA extraction [19–21].

Also, in some cases where human remains are found and no relatives have been identified for genetic identification, forensic anthropology tries to provide information regarding the gender and height of the deceased and helps in estimating the time since death and the cause of death (if the remains provide relevant evidence). Approximate height can be determined by looking at the measurements of the bones; the best way to find approximate height is to measure the femur bone.

In forensic anthropology, the preferred method for establishing the identity of skeletal remains is dental identification. In some cases, when the teeth or skull may be missing, other alternatives to dental identification may be used.

Advancements in the field of forensic genetics through automate systems have aided in reducing the time that is spent on obtaining genetic profiles. Also, the introduction of next-generation system (NGS) and massive parallel sequencing (MPS) systems has enabled the concomitant analysis of multiple STR, and SNPs of human remains to identify the person [22–26].

To our knowledge, this is the first forensic laboratory in Romania regarding forensic human DNA identification from bones and tooth, where liquid nitrogen was not used with an automate protocol and DNA extraction from skeletonized remains was performed within three hours.

#### **5. Conclusion**

The biological material subjected to DNA identification analyzes in forensic medicine is in different taphonomy conditions and the selection of appropriate working samples is crucial.

In genetic human identification, automate system for DNA extraction from different biological samples is better than manual extraction, which is time consuming and presents the risk of errors.
