Extraction of mitochondrial DNA from tooth dentin: application of two techniques
- Author:
Ahmad Azlina a,b*
;
Berahim Zurairah a
;
Sidek Mohamad Ros b
;
Mokhtar Khairani Idah a
;
Samsudin Abdul Rani c
- Publication Type:Journal Article
- Keywords:
Dentin, extraction, mitochondrial DNA, tooth.
- From:Archives of Orofacial Sciences
2011;6(1):9-14
- CountryMalaysia
- Language:English
-
Abstract:
Mitochondrial DNA (mtDNA) is a hereditary material
located in mitochondria and is normally maternally inherited.
Mutational analysis performed on mtDNA proved that the
mutations are closely related with a number of genetic
illnesses, besides being exploitable for forensic identification.
Those findings imply the importance of mtDNA in the scientific
field. MtDNA can be found in abundance in tooth dentin where
it is 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. Teeth used for the study was collected from Dental
Clinic, Hospital Universiti Sains Malaysia. After the removal of
tooth from the tooth socket of the patient, the tooth was kept at
-20C until use. Later, pulp tissue and enamel was excised
using dental bur and only the root dentin was utilized for the
isolation of mtDNA by crushing it mechanically into powdered
form. MtDNA was extracted using the two published methods,
Pfeifer and Budowle and then subjected to spectrophotometry
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 capable for the extraction of mtDNA from tooth
dentin.
- Full text:W020150629335166796295.pdf
