Ahmad, Azlina
(2011)
Comparison of DNA extraction methods from dentine (of tooth) as a source of genetic material.
Comparison of DNA extraction methods from dentine (of tooth) as a source of genetic material.
Abstract
Mitochondrial DNA (mtDNA) is a maternally inherited hereditary material locatt-d in
mitochondria. Previous studies on mtDNA proved that the mutations are closely related
with a number of genetic illnesses and exploitable for forensic identification. Those
findings imply the importance of mtDNA in the scientific field. The mtDNA can be
found in abundance in tooth dentin where it is being kept protected by the enamel, the
hardest outer part of the tooth. In this study, two techniques of mtDNA extraction were
compared to determine the efficacy between the two techniques. After the extraction of
tooth, the tooth was kept at -20°C until use. Later, pulp tissue and enamel were excised
out using dental bur and only the root dentin was utilized for the isolation of mtDNA by
crushing it mechanically into powder form. The mtDNA was then extracted using the two
published methods, Pfeiffer and Budowle and then subjected to spectrophotometry for
DNA quantification and purity, Polymerase chain reaction (PCR) amplification of
hypervariable-two region of mtDNA, followed by DNA sequencing to analyze the
reliability of the extraction techniques. In conclusion, both techniques proved to be
efficient and consistent for the extraction of mtDNA from tooth dentin.
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