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Genomics and Applied Biology 2015, Vol. 6, No. 1, 1-10
http://gab.biopublisher.ca
8
Polyacrylamide Gel Electrophoresis systems were
illuminated from above using and white fluorescent
light source was given in Figure 5. In this study, DNA
based sex determination test was used to aid in the
verification of individual identification in order to
compare to historical documentation of burials.
Applying sex determination test based on primers
specific to human amelogenin on the short arm of both
the X (AMELX) and Y (AMELY) chromosomes on
genomic DNA from males and females. Therefore,
single copy X-Y homolog related amelogenin was
amplified under optimal PCR condition. The PCR
amplification from DNA of different ancient bones in
male and female using AMEL primers was displayed a
sex-spesific band pattern showing a length variation
characteristic for AMELX and AMELY. Female
related to AMELY showed a single 106 bp band
whereas male an additional band of 112 bp was
observed from 50 bones (Figure 4). However, DNA
from 10 bones didn’t amplify the AMEL fragments
due to contamination of DNA during preservation of
bone on buriel. The typing results for the amelogenin
locus are given in Figure 4 and 5. With the exception
of fifty samples positive results could be obtained for
all bones leading to a success rate of 90%. Ancient
DNA (aDNA) and proteins provide valuable clues to
questions about nutrition, domestication, population
genetics, kinship reconstruction and human evolution.
By investigating ancient biomolecules with the use of
newer molecular biology techniques and robust
procedures of inference genetic data from both
archaeological remains and living populations,
molecular anthropology has begun to draw more
informed conclusions about human evolutionary
history. Ancient DNA can shed light on the
relationships between populations and how they
dispersed through the ancient world and validate
evolutionary hypotheses inferred from archaeological,
linguistic and historical records. Also, aDNA can help
solve archaeological puzzles and build up a picture of
the demography of past societies by identifying the
sex of skeletons that cannot be determined by
osteology and to assess the degree of maternal
relatedness in multiple burials. The remarkable thing
about sexual differentiation is its diversity. That males
are the heterogametic sex, larger
than females, more
aggressive than females, and the ‘non-default’ mode
of sexual differentiation
are concepts not valid
throughout most of the
animal kingdom. Sex
chromosomes are characteristic only of land animals.
In birds, the heterogametic sex
is female and the sex
chromosomes are not related
to those of mammals.
External factors such as temperature determine sex in
lower vertebrates, and there is no similarity
among
sex-determining genes of different species.
3 Concluding Remarks
Ancient DNA has been proven to be useful genetic
markers for studies of kinship, parentage, gene
mapping and sex determination. They are highly
polymorphic, easily scored, single-locus markers that
are readily applied to small or degraded DNA samples,
including museum specimens (Ellegren, H., 1991).
Genetic typing of ancient bones means trying to
obtain multilocus STR profiles from highly degraded,
minimal, or even no detectable amounts of human
DNA. Additionally, there are several other factors
hampering a successful amplification, e.g. burial
conditions such as temperature, humidity, and huminic
acids. Our results were according to those findings
since there was absolutely no correlation between the
age of the bone and the amplification success.
Therefore, our idea was to create an experimental
approach to systematically investigate the age and
environmentally caused degradation of DNA. Each 1
ml of DNA extract was employed to amplify 15 STR
loci plus amelogenin sex determination
simultaneously. The artificial aging process led to an
amplification pattern which is supposed to be typical
for aDNA with weaker signals or total allelic drop
outs for the longer PCR products as described in Ref.
Burger J., et.al., (1999), thus providing a hint for
simulating the original degradation process as it
occurs in bone tissues and for amplification of
authentic aDNA from historical bones since the typing
patterns were similar. The present method to
determine sex employs PCR and a nonradioactive dot
blot procedure to examine sequences from the
amelogenin gene on the X and Y chromosomes (Herr
C. M., et. al., 1990). One ancient individual was
classified as a male based on morphology, but the
PCR products hybridized only with the X specific
