Forensic Science International 353 (2023) 111882

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Forensic Science International

journal homepage: www.elsevier.com/locate/forsciint

Comparative analysis of DNA preservation in permanent and deciduous

teeth of adults and non-adults: Implications for archaeological and
forensic research
Tamara Leskovar a, 1, Irena Zupanič Pajnič b, *, 2
a
Centre for Interdisciplinary Research in Archaeology, Department of Archaeology, Faculty of Arts, University of Ljubljana, Ljubljana, Slovenia
b
Institute of Forensic Medicine, Faculty of Medicine, University of Ljubljana, Korytkova 2, 1000 Ljubljana, Slovenia

A R T I C L E I N F O A B S T R A C T

Keywords: This study investigates the preservation of DNA in different categories of teeth, including permanent and de­
DNA preservation ciduous, fully developed and not fully developed, in both adults and non-adults. Teeth were sampled from a
Tooth type modern-era cemetery in Ljubljana, Slovenia. DNA extraction was performed using a full demineralisation pro­
Permanent teeth
tocol. DNA quantity and quality were assessed using qPCR analyses, and autosomal STR typing was conducted to
Deciduous teeth
Skeletonized remains
verify genetic profiles. Results revealed significant differences in DNA preservation among various tooth cate­
gories. Fully developed permanent teeth of adults exhibited the highest DNA yields, attributed to their fully
developed roots and thicker cementum, which is rich in DNA. Deciduous teeth, with thinner enamel and
cementum, showed lower DNA preservation regardless of developmental stage. Non-adult teeth generally yielded
less DNA compared to adults, even when considering only fully developed permanent teeth, indicating factors
beyond developmental stage. These findings suggest that, in archaeological and forensic contexts, researchers
should prioritize fully developed permanent teeth for DNA analysis due to their superior preservation. Addi­
tionally, this study underscores the importance of considering tooth type and developmental stage when
selecting samples for genetic analysis in cases where petrous bone is unavailable, expanding our understanding of
DNA preservation in human remains.

1. Introduction
Recovery and analyses of DNA have become an effective and rela­
tively accessible way to extract information from human skeletonized
remains in archaeological and forensic cases. Nowadays, even analyses
of DNA from highly degraded skeletal tissues are possible, allowing re­
searchers to extract unique genetic information. Though research shows
that petrous bone is optimal skeletal element to use for DNA analyses
[1–8], it is not always available for sampling. Other bones of the skel­
eton do show great potential [9–12], yet teeth, if available, seem to be
the second choice [7,8,13,14]. Teeth are preferred due to their hard
mineral composition and low porosity, which makes them less prone to
taphonomy and contamination when compared to bones [15–17].
Human teeth can be separated into different categories. Based on
development, teeth can be deciduous (milk teeth) or permanent.
Deciduous teeth are further divided into incisors, canines and molars,
while permanent teeth also have premolars [18]. Deciduous teeth start
to form at approximately 30 weeks in-utero and finish their develop­
ment with all the roots closed at the age 3.5 years ± 1 year. Permanent
teeth start to develop at the age of 4.5 months ± 3 months and finish
their development with the third molars roots closed at around the age
of 23 years [19]. As teeth differ in developmental stage or serve different
functions, different teeth have different structural properties. Incisors
are flat and have one root, canine are more conical, with one root, which
is longer in comparison to other teeth, premolars are round with two
cusps and are usually single rooted, but can have two roots, while molars
are larger, squarer and have more cusps and roots than other teeth [18].
Deciduous teeth have thinner and shorter roots and thinner dentine
layer and enamel when compared to permanent teeth. On the other
hand, deciduous teeth have larger pulp chamber with blood vessels and

* Corresponding author.
E-mail address: irena.zupanic@mf.uni-lj.si (I.Z. Pajnič).
1
Orcid ID: 0000-0002-4585-4726
2
Orcid ID: 0000-0002-6704-015X

https://doi.org/10.1016/j.forsciint.2023.111882
Received 6 October 2023; Received in revised form 30 October 2023; Accepted 7 November 2023
Available online 10 November 2023
0379-0738/© 2023 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
T. Leskovar and I.Z. Pajnič
nerves, than permanent teeth [18]. Furthermore, due to posteruptive
maturation, enamel becomes more mineralized with age and thus less
prone to decay, especially shortly after eruption [20], but likely loses
this advantage with aging. Studies indicate that properties such as
dental elasticity, durability, and fracture resistance decrease with age
[21–23], making teeth more prone to deterioration [24].
When sampling teeth for DNA analyses, is it advised to choose least
damaged teeth with intact root systems, as in such cases DNA has highest
rate of survival [14,25]. Additionally, molars are recommended, as they
are multi-rooted with large pulp chamber [26–29]. However, molars are
not always available and Heathfield et al. [30] showed that there are no
significant differences in the preservation of DNA in the four types of
teeth (incisors, canine, premolars and molars), indicating that sampling
could be broadened to other tooth types. In general, DNA analyses
performed on teeth focus on pulp tissue, which degrades relatively
quickly and is thus not necessarily the best source in case of tapho­
nomically altered skeletonized remains [31,32]. Consequently, sam­
pling cementum presents an alternative for DNA analyses, as it is more
resistant to degradation [14,31,33].
Despite being included in various archaeological and forensic
research [25,34–39], there is no specific data or guidelines on the DNA
analyses performed on teeth of non-adults or on deciduous teeth.
Though for DNA analyses sampling tooth cementum, preferably of fully
developed teeth with closed, undamaged roots is advised, this is not
always an option. In case of non-adults, permanent teeth might still be
developing or are not even present. Thus, deciduous teeth or partially
developed teeth are the only option. Furthermore, focusing on one part
of the tooth might also not be possible as deciduous and still developing
teeth are usually small and light, and the whole tooth needs to be used
for the analysis. The aim of this study is to start filing these gaps in
knowledge by comparing DNA preservation of permanent and decidu­
ous, fully developed and not fully developed teeth of adults and
non-adults.
2. Materials and methods
2.1. Sample selection
An archaeological excavation of a modern-era cemetery that was
active between the early 16th and late 19th centuries in Ljubljana – Polje
took place in 2020, and 216 graves with one third of adult and two thirds
of non-adult skeletons were uncovered [40]. For the analysis, 52 teeth
from 15 non-adults and 11 adults were sampled and one to three (mostly
two) teeth were sampled per skeleton (Supplementary material Table 1).
From non-adults, 16 deciduous (11 molars and 5 premolars) and 17
permanent teeth (12 molars and 5 canines) were collected, and from
adults 19 permanent teeth (8 molars, 2 premolars and 9 canines) were
collected. Out of 36 permanent teeth, 14 were not fully developed
(partially developed rooted or open apex). Pathology was observed on
11 teeth, in one case the crown had a malformation and in ten cases mild
carious lesions were observed.
2.2. DNA extraction
DNA extraction was performed according to the protocol of Zupanič
Pajnič [41]. Because extraction method is optimized for 0.5 g of
bone/tooth powder and deciduous and still developing teeth are small
and light, whole teeth were grounded for non-adults. Whole teeth were
used for the analysis from adults as well so that comparison between
adult and non-adult teeth was possible. Each tooth was chemically
cleaned (5% Alconox detergent (Sigma-Aldrich, St. Louis, MO, USA),
water and 80% ethanol) and UV irradiated for 30 min using
BLX-Multichannel BioLink DNA Crosslinker (Vilber, Collégien, France)
to remove surface contamination and grounded into a fine powder
using a Bead Beater MillMix 20 homogenizer (Tehtnica, Domel,
Železniki, Slovenia) for 1 min at a frequency of 30 Hz. The metal vials
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Forensic Science International 353 (2023) 111882
of the homogenizer and teeth were cooled with liquid nitrogen to
prevent overheating. Before use, metal vials and metal balls were
cleaned with 6% sodium hypochlorite, sterile bi-distillated water, and
80% ethanol, followed by sterilization in a Europa B xp sterilizer
(Tecno-Gaz, Parma, Italy), and 30 min of UV irradiation using
BLX-Multichannel BioLink DNA Crosslinker (Vilber). From each tooth,
0.5 g of tooth powder was used for extraction. To prevent and follow
possible contamination with modern DNA, special measures [42,43]
were followed, extraction-negative controls (ENC) were added to each
batch of samples [44], and all persons involved in analyses were
include in an elimination database (their saliva was collected with
cotton swabs, and extracted DNA was used for STR typing). Tooth
sample purification was carried out with the EZ1 Advanced XL ma­
chine (Qiagen). Instructions provided by the manufacturer [45] were
followed for using the EZ1&EZ2 DNA Investigator kit (Qiagen) for
processing buccal swabs, and this was also in line with the previously
published protocol [41] for processing tooth samples.
2.3. DNA quantification
To determine the concentration of DNA and its degradation in teeth
extracts, qPCR analyses were performed amplifying short (Auto target)
and long (Deg target) autosomal targets using the PowerQuant System
(Promega). DNA quantity was determined from Auto target measures,
and the degradation index was calculated from the Auto/Deg ratio. We
used an internal PCR control (IPC shift) to detect any possible presence of
PCR inhibitors. The Y-chromosomal target included in the same kit was
used to detect the presence of male DNA. Analyses were performed
following the technical manual [46] with the QuantStudio 5 Real-Time
PCR system, the PowerQuant Analysis Tool (https://worldwide.prom­
ega.com/resources/tools/powerquant-analysis-tool/), and Quant-Studio
Design and Analysis Software 1. 5. 1 (Applied Biosystems, AB, Foster
City, CA, USA).
The threshold for the Auto/Deg ratio was set at 2 and the IPC shift
value was set at 0.30, as recommended by the manufacturer [46].
Laboratory plastic and reagent cleanliness was verified through analysis
of negative template controls and extraction-negative controls. Tooth
extracts, positive controls, and negative controls were quantified in
duplicate, following the manufacturer’s recommendations [46]). For all
of the tooth samples, the DNA quantity obtained from 1 g of tooth was
calculated on the basis of the Auto target, and this was expressed in ng
DNA/g of tooth.
2.4. Autosomal STR typing
The autosomal STR typing kit PowerPlex ESI 17 Fast System
(Promega) was used for STR typing. PCR was performed following the
recommendations of the manufacturer [47], using the Nexus Master­
Cycler (Eppendorf, Hamburg, Germany). Amplification included both
positive and negative PCR controls. Variation in the final volume of
tooth extracts used in the PCR reaction was based on the quantification
results of the PowerQuant Auto target. If the quantification was 0.058 ng
per μl of extract or higher, 1 ng of DNA was used as a template. The
maximum DNA extract volume (17.5 μl) was used if the quantification
was lower than 0.058 ng per μl of extract. Maximum volume was also
used for amplification of ENCs. Genetic profiles for the samples were
acquired using the SeqStudio Genetic Analyzer for HID (Thermo Fisher
Scientific, TFS) combined with the WEN Internal Lane Standard 500
(Promega), SeqStudio Data Collection Software v 1.2.1 (TFS), and
GeneMapper ID-X Software v 1.6 (TFS). All ENC samples employed to
monitor possible contamination with modern DNA were also typed, and
STR typing of elimination database samples was performed to check the
authenticity of isolated DNA and to exclude modern DNA contamination
of extracts.
T. Leskovar and I.Z. Pajnič
2.5. Statistical analysis
According to the research problem, comparisons were made between
different classes of teeth:
– between the teeth of adults and non-adults,
– between deciduous and permanent teeth,
– between full developed and not fully developed permanent teeth,
– between fully developed and not fully developed deciduous teeth,
– between one rooted or more rooted permanent teeth,
– between one rooted or more rooted deciduous teeth.
According to the obtained results and in search of their interpreta­
tion, additional comparisons were made accordingly. In each case, the
amount of DNA (expressed in ng DNA/g of tooth), the degradation ratio
(Auto/Deg ratio of the DNA) and STR typing results were compared.
The normality and homogeneity of variance was tested using the
Kolmogorov–Smirnov test (with Lilliefors significance correction).
Following the recommendations for statistical analyses in medical
studies [48–50], the research hypotheses were tested using the 95%
confidence intervals for means or medians. For the analyses, computer
program IBM SPSS Statistics for Windows, version 25.0 (Statistical
Package for the Social Sciences Inc., Chicago, IL, USA) was used. As
sample sizes are relatively small, confidence intervals can have limited
power to detect significant differences [48]. Thus, formulated hypoth­
eses were also tested using p values. Significance was set as p ≤ 0.05.
Data was obtained from teeth of 15 non-adults and 11 adults,
together from 52 teeth. In seven cases no data was obtained for neither
amount of DNA (expressed in ng DNA/g of tooth) and Auto/Deg ratio. In
all seven cases the teeth were from non-adults. Since there was not
enough data for these seven cases, they were omitted from the com­
parisons. There were also 13 cases in which the amount of DNA was
obtained while the value for the Auto/Deg ratio was not possible to
calculate because no Deg targets were amplified. The Auto/Deg ratio for
these 13 cases was calculated using the following formula: value = Max
+ SD. Missing values of the degradation ratio were set to 558.64, as max
value was 472.94 and SD was 85.70. STR typing was successful in 17
cases, in each case on a permanent tooth. Thus, comparisons were only
possible between not fully developed and fully developed permanent
teeth of adults and non-adults.
The normality of the distribution of amount of DNA, Auto/Deg ratio
and STR typing was checked, using the Kolmogorov-Smirnov test
(Supplementary material Statistics). DNA and Auto/Deg ratio were not
normally distributed (sig. < 0.05), thus non-parametric tests and me­
dians were used to test the formulated research hypotheses. STR typing
was normally distributed, thus parametric tests and means were used for
additional comparisons.
3. Results
Results are gathered in Supplementary material Table 2.
3.1. DNA quantification
Supplementary material Table 2 presents tooth sample characteris­
tics and summarizes the PowerQuant (Promega) results for DNA quan­
tity and quality (Auto and Deg in ng DNA/μl of extract, Auto/Deg ratio
and the amount of DNA obtained from 1 g of tooth in ng of DNA. No DNA
was obtained from seven teeth (all were from non-adults) and less than
0.5 pg of human DNA per μl of extract was detected in 15 tooth samples
(12 from non-adults and three from adults). It was not possible to
determine the Auto/Deg ratio for 20 samples (18 non-adult and 2 adult
teeth). Extracted tooth DNA was slightly to severely degraded with
degradation index ranged between 1 and 473. An IPC shift value was
lower than 0.3 in all extracts and no amplicons were detected for ENC
samples (data not shown).
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Forensic Science International 353 (2023) 111882
3.2. STR typing
All samples were typed for autosomal STRs, and only 17 of them
produced useful genetic profiles, among them only four tooth samples
originated from non-adults (see Supplementary material Table 2).
Thirteen samples produced STR profiles with 12 or more STRs amplified
and four of them generated profiles with 7–11 successfully amplified
STRs. For all the skeletons used in this study, petrous bones, tali,
calcanei and femurs were analyzed in our previous study and consensus
profiles were generated [40]. DNA profiles obtained from tooth extracts
matched consensus profiles of corresponding skeletons, indicating
endogenous tooth DNA and together with clean ENCs (ENC samples did
not produce profiles), and no match with elimination database, elimi­
nates contamination issue. For the most of teeth from non-adult skele­
tons, no DNA or low-quantity DNA was extracted and consequently,
autosomal STR typing failed, showing less DNA yield in teeth sampled
from non-adult skeletons than from adult skeletons.
SM Table 1–6 show genetic profiles obtained from different amounts
of DNA and from DNA samples with different degradation rates. SM
Figs. 1–3 show genetic profiles of tooth samples from which higher DNA
yields were obtained and one ng (SM Figs. 1 and 2) and 452 pg of DNA
were amplified (SM Fig. 3). Because samples with different degradation
indexes were amplified, full profile was obtained only from sample 238
(SM Fig. 1) with degradation index of 10.94. In sample 209 (SM Fig. 2)
partial profile with one locus drop-out was obtained even if 1 ng of DNA
was used for PCR because of higher degradation of sample 209 (degra­
dation index of 92.25 was measured). Sample 171 produced partial
profile with several loci and allelic drop-outs (see SM Fig. 3) indicating
high DNA degradation (degradation index of 168.84 was calculated).
SM Fig. 4–6 show genetic profiles of tooth samples from which lower
DNA yields were obtained and 47 pg (SM Fig. 4), 54 pg (SM Fig. 5) and
39 pg of DNA were amplified (SM Fig. 6). Because samples with different
degradation indexes were amplified, full profile was obtained only from
sample 262 (SM Fig. 4) where degradation index was very low (3.09). In
sample 228 (SM Fig. 5) and sample 123 (SM Fig. 6) only partial profiles
with several loci and allelic drop-outs were generated according to
higher degradation of DNA (for sample 228 degradation index was 8.58,
and for sample 123 it was 12.32). However, degradation was not so high
as in samples 209 and 171 and partial profiles were generated despite
very low DNA input.
3.3. Statistics
There were no significant differences in case of degradation ratio
when comparing different classes of teeth. Significant differences were
obtained when comparing the amount of DNA in the teeth of adults and
non-adults, in deciduous and permanent teeth, and in fully developed
and not fully developed deciduous teeth. There were no significant
differences between deciduous or permanent one rooted or more rooted
teeth (Supplementary material Statistics).
Results suggest significant differences (p = 0.002) among non-adults
and adults in the amount of the DNA extracted from teeth. There is more
DNA extracted from the teeth of adults (Me=1.3, SE=0.47) than of non-
adults (Me=0.18, SE=0.10) (Fig. 1).
Significant differences (p = 0.011) were also observed among de­
ciduous and permanent teeth in the amount of the DNA. There is more
DNA extracted from the permanent teeth (Me=0.89, SE=0.30) than
from deciduous teeth (Me=0.06, SE=0.12) (Fig. 2).
The difference in the amount of the DNA in not fully developed and
fully developed permanent teeth is statistically significant (p = 0.008).
There is more DNA extracted from the fully developed permanent teeth
(Me=1.14, SE=0.41) than from the not fully developed permanent teeth
(Me=0.34, SE=0.25). However, the confidence interval shows over­
lapping and statistical significance is only seen in p-value (Fig. 3; Sup­
plementary material Statistics).
Results suggest significant differences (p = 0.030) among non-adults
T. Leskovar and I.Z. Pajnič Forensic Science International 353 (2023) 111882

Fig. 1. 95% confidence intervals for medians for the amount of the DNA extracted from teeth (for non-adults and adults, separately).

Fig. 2. 95% confidence intervals for medians for the amount of the DNA extracted from teeth (for permanent and deciduous teeth, separately).

and adults in the STR typing. More STRs were amplified from teeth of
adults (Me=13.5, SE=0.54) than of non-adults (Me=10.5, SE=1.4).
However, the confidence interval shows overlapping and statistical
significance is only seen in p-value (Fig. 4; Supplementary material
Statistics).
4. Discussion
Significant differences in the amount of DNA were observed between
teeth of adults and non-adults, between deciduous and permanent teeth
and between fully developed and not fully developed permanent teeth.
Permanent, fully developed teeth of adults presented highest amounts of
DNA. As previous research has shown that damaged or open roots fasten
the degradation of DNA [14,25], better preservation of DNA in fully
developed teeth is completely understandable. Though, this was only
observed in permanent teeth, while not in deciduous. The reason could
be low number of not fully developed deciduous teeth (only two).
Another explanation is the also observed difference in permanent and
deciduous teeth, as the former presented significantly higher amounts of
DNA. In comparison to permanent teeth, deciduous teeth are more prone
to degradation regardless of their developmental stage, as they have less
mineralized, thinner enamel and thinner cementum [18,51], leading to
faster degradation. This can be observed in degradation ratio, though
not significant, double in deciduous teeth (310,9 ± 267.7) when
compared to permanent teeth (144.9 ± 216.1). The logic conclusions
would thus be, that teeth of non-adults presented lower quantity of DNA
because there are deciduous teeth present in the non-adult group.
However, when only comparing permanent, fully developed teeth of
adults and non-adults, the difference remained significant (p = 0.016),
with average amount of DNA/g of tooth 0.13 ± 0.09 in non-adult teeth

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T. Leskovar and I.Z. Pajnič Forensic Science International 353 (2023) 111882

Fig. 3. 95% confidence intervals for medians for the amount of the DNA extracted from permanent teeth (for not fully developed and fully developed
teeth, separately).

Fig. 4. 95% confidence intervals for STRs (for teeth of adults and non-adults, separately).

and 1.45 ± 2.03 in adult teeth. The significant difference remained
present even when comparing only permanent molars, as adult molars
presented on average 3.06 ± 2.7 amount of DNA/g of tooth, while non
adult molars 0.13 ± 0.09 amount of DNA/g of tooth. Caution is advised
as with such data fragmentation sample number is low, yet it indicates
there is some other reason for the better preservation of DNA in teeth of
adults than developmental stage or type of tooth. STR typing results
further confirm this, as the only significant differences were observed
between the teeth of adults and non-adults (p = 0.030), while non be­
tween fully developed and not fully developed teeth (see Supplementary
Material Statistics, Appendixes 7 and 8). Degradation ratio presented no
significant differences between the fully developed permanent teeth of
adults and non-adults, but it should be highlighted that on average, the
ratio was almost double in non-adults (209.2 ± 248.4) when compared
to adults (114.9 ± 189.1). One possibility is differences in enamel
mineralization, as highly mineralized enamel offers protection for DNA
[16,52]. Studies suggest enamel mineralization is highest in first couple
of years after eruption and decreases with old age [20]. Analysed fully
developed first permanent molars of non-adults in this study originate
from children aged 11.5 – 13.5 years. As first permanent molars erupt
around the age of 6 years, the enamel should already be highly miner­
alized. Thus, enamel maturity might also not be influencing DNA pres­
ervation in our case. A more thorough analysis revealed either an
enamel malformation or presence of carious lesions on all the analysed
non-adult permanent, fully developed teeth. Comparison of amount of
DNA in all the permanent teeth with (ngDNA/g tooth = 1.13 ± 1.84) or
without pathology (ngDNA/g tooth = 1.26 ± 1.95) presented no sig­
nificant differences, which agrees with other studies [53]. Thus,

5
T. Leskovar and I.Z. Pajnič
pathology is also not a tangible reason for the differences. Especially, as
observed carious lesions were relatively small and superficial, not
affecting the pulp, and the degradation ratio was lower in teeth with
pathology (68.5 ± 163.7) when compared to teeth without pathology
(182.88 ± 216.40). As the whole teeth were used for the analysis,
including DNA rich tooth cementum, this is the most likely reason for the
observed differences between adult and non-adult fully developed teeth.
Tooth cementum is rich in mtDNA and nuclear DNA [14,16,54–57] and
contrary to enamel, cementum apposition continues throughout life
[58]. Thus, the thickness of cementum increases with age [59] and could
cause differences between non-adult and adult permanent teeth. This
would also further support the differences between permanent and de­
ciduous teeth, and fully- and not-fully developed teeth. Though these
differences could be attributed to taphonomy with deciduous/not-fully
developed teeth being more susceptive to taphonomic factors, they
also have shorter/not yet fully developed roots and thinner cementum
[18,51,60], which could result in lower quantities of DNA. A reliable
answer could be obtained through a direct comparison of tooth
cementum of teeth of adults and non-adults. However, some of the
analysed teeth of non-adults were not fully developed, having incom­
plete roots. As cementum forms on the roots, these not fully developed
teeth could distort the comparisons. To avoid this, whole tooth was used
for the DNA analyses in this study.
5. Conclusions
1. Tooth Type Matters: This study highlights the significance of tooth
type in DNA preservation. Permanent teeth, with their thicker
cementum and fully developed roots, consistently outperformed decid­
uous teeth in terms of DNA yield. Researchers should prioritize the use of
permanent teeth when available for genetic analysis, as they offer a
more reliable source of DNA, especially in challenging archaeological
and forensic cases.
2. Age-Related Variations: The study demonstrates that non-adult
teeth generally yielded lower quantities of DNA compared to adult
teeth. Even when considering only fully developed permanent teeth,
adults consistently provided higher DNA yields. This suggests that age-
related factors, beyond tooth developmental stage, influence DNA
preservation. Further research is needed to explore these factors and
their impact on DNA preservation in non-adult individuals.
3. Cementum as a DNA Source: Tooth cementum emerges as a
valuable alternative for DNA analysis, given its resistance to degrada­
tion. The thickness of cementum increases with age, potentially
contributing to higher DNA yields in adult teeth. Researchers should
consider tooth cementum as a viable source of genetic material, espe­
cially in cases where other tooth components may be compromised.
4. Implications for Archaeological and Forensic Research: These
findings have practical implications for both archaeological and forensic
investigations. Selecting the most suitable teeth for DNA analysis can
enhance the chances of obtaining reliable genetic information from
skeletonized remains. Understanding the factors that influence DNA
preservation in teeth can lead to more informed sampling strategies and
improved success rates in genetic analyses.
This study contributes valuable insights into DNA preservation in
different categories of teeth and emphasizes the importance of tooth
type and age in genetic analyses of archaeological and forensic samples.
These findings offer guidance for researchers in optimizing their sam­
pling strategies and maximizing the success of DNA extraction from
human skeletal remains.
Ethical approval
This research project was approved by the Medical Ethics Committee
of the Republic of Slovenia (0120-526/2021/7).
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Forensic Science International 353 (2023) 111882
Funding
The work was supported by Slovenian Research and Innovation
Agency: project J3-3080 and Research Group Archaeology, 0581-012).
CRediT authorship contribution statement
Tamara Leskovar: Funding acquisition, Investigation, Methodol­
ogy, Software, Visualization, Writing – original draft. Irena Zupanič
Pajnič: Conceptualization, Formal analysis, Funding acquisition,
Methodology, Supervision, Validation, Writing – review & editing.
Declarations
Research involving human participants and/or animals Research
involves ancient skeletons and genetic profiles of persons included in
elimination database and from them informed consents were obtained
and submitted to the Medical Ethics Committee of the Republic of
Slovenia. After submission, the Medical Ethics Committee of the Re­
public of Slovenia approved the research (number of approval is 0120-
526/2021/7.
Informed consent
This research project was approved by the Medical Ethics Committee
of the Republic of Slovenia (0120-526/2021/7) and informed consents
of persons included in elimination database were submitted to the
Medical Ethics Committee of the Republic of Slovenia.
Declaration of Competing Interest
The authors declare no conflict of interest.
Data availability
The authors declare that all the data are available upon reasonable
request.
Acknowledgements
This study was financially supported by the Slovenian Research
Agency (the project “Inferring ancestry from DNA for human identifi­
cation” J3-3080). The authors would like to thank The Ljubljana
Museum and Galleries (MGML) and the responsible curator Martin
Horvat for including the archaeological human remains from the
museum into our study. Authors are grateful to Teo Mlinšek and Tadej
Počivavšek for DNA extraction.
Appendix A. Supporting information
Supplementary data associated with this article can be found in the
online version at doi:10.1016/j.forsciint.2023.111882.
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