![]()
Genomics and Applied Biology 2015, Vol. 6, No. 1, 1-10
http://gab.biopublisher.ca
9
oligonucleotide. This may be the result of insufficient
DNA, or mutations in the priming or oligonucleotide
binding sites on the Y chromosome copy of the
amelogenin gene. Another molecular technique for sex
determination that has been applied to skeletal
remains was developed by Sullivan et al. (1993).
For
this method, which also uses the amelogenin gene, a
short DNA fragment from intron 1
is amplified that
contains a
6
bp deletion in the X chromosome
sequence that is not present in the corresponding Y
chromosome sequence. Gill et al. (1994)
employed
this technique to determine the sex of skeletal remains
found near Ekaterinburg, Russia, that were reputed to
be those of the Romanovs, their doctor, and three
servants. The method described in this paper, like the
technique designed by Sullivan et al. (1993),
solves
the problem encountered when using the Y repeat
sequences. Our method, however, appears to be more
sensitive to very low quantities of DNA, such as those
likely to be found in ancient bones. A dilution series
testing the sensitivity of the primers indicates that they
can begin amplification from as few as one or two
copies of the gene
.
Sex determination using DNA can
be valuable for both forensic and archaeological
research. Standard osteological methods, however, are
less expensive and more rapid when the skeletons of
adults are complete and the bones are in good shape.
For archaeological research, the use of DNA to
determine the sex of juveniles provides an opportunity
to extend traditional mortuary analyses through the
inclusion of children of known sex. Molecular
analyses can also address questions regarding the sex
of adult skeletons that fall in the overlapping range of
male and female morphological variation. By using
this method in combination with routine genotyping
more information about a material under investigation
can be obtained. In addition, the amplification of the
AMEL gene can also be used as an internal control.
In conclusion our findings show that the PCR assay
based on the AMEL gene is reliable for sex
identification of fossil bone remains in Koranza and
Necropal area are situated in the region of modern city
Mugla in Turkey. The advantage of this assay is that
neither additional control amplicons with a second
locus specific autosomal primer pair nor restriction
endonuclease steps are necessary for sex
determination and control of the PCR reaction.
However, despite these objections and characteristic
features of aDNA mentioned above, it can be shown
that the molecular approach is the most powerful tool
for the identification and reconstruction of kinship of
skeletal human remains of archaeological excavations.
These validated protocols allow the assignment of
unknown men to every major branch of the global
human population. Hopefully these protocols will
encourage new research groups to implement a
broader range of anthropological surveys,
archaeological excavations and archaeometry studies
etc. Furthermore, there is not the only parameter that
determines the overall specificity and sensitivity of the
PCR reaction; primer design and optimization of PCR
parameters also have a profound effect. Results of the
present our work demonstrate that the primers utilised
in this test (Amel A and Amel B) provide robust and
highly efficient amplification. It is envisaged that this
test will prove to be an advantageous addition to other
methods of forensic DNA analysis. The size difference
between the amplified segments of X and Y copy of
AMG was not big enough to be detected clearly on
agarose or polyacrylamide gel electrophoresis (PAGE).
For that reason, we searched the list of commercial
restriction enzymes and find a new enzyme capable of
recognizing and cleaving the PCR product for Y copy of
AMG, but not the X copy. The molecular determination
of gender based on AMG PCR/Restriction enzyme
digestion was compared with anthropometric reports. At
the beginning stages of the project the molecular sex
determination was both different from anthropometric
reports and also not reproducible. However, after
optimizing the procedure and setting guidelines to
eliminate the risk of contamination we were able to have
reliable and reproducible molecular sex determination.
According to study, 36 female from 100 samples, 10 of
them do not proliferate in the rest of the samples are
composed of male subjects.
Acknowledgment
Samples were kindly provided by Selcuk University, Arts and
Science Faculty, Department of Archeology. I also thank
Professor A. Ahmet TIRPAN and his fellowship and were
studied experimental step related to research in my
Archaeometry and Biotechnology Laboratory.
