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Comparison of three methods of DNA extraction from human bones with different degrees of degradation
Authors:Jakubowska Joanna  Maciejewska Agnieszka  Pawłowski Ryszard
Affiliation:(1) Intercollegiate Faculty of Biotechnology of University of Gdańsk and Medical University of Gdańsk, Kładki 24, 80–822 Gdańsk, Poland;(2) Institute of Forensic Medicine, Medical University of Gdańsk, Dębowa 24, 80–204 Gdańsk, Poland;(3) Institute of Forensic Research, Westerplatte 9, 31–033 Krak?w, Poland;
Abstract:There is a necessity for deceased identification as a result of many accidents and sometimes bones are the only accessible source of DNA. So far, a universal method that allows for extraction of DNA from materials at different stages of degradation does not exist. The aims of this study were: the comparison of three methods of DNA extraction from bones with different degree of degradation and an evaluation of the usefulness of these methods in forensic genetics. The efficiency of DNA extraction, the degree of extract contamination by polymerase chain reaction (PCR) inhibitors and the possibility of determining the STR loci profile were especially being compared. Nuclear DNA from bones at different states of degradation was isolated using three methods: classical, organic phenol–chloroform extraction, DNA extraction from crystal aggregates and extraction by total demineralisation. Total demineralisation is the best method for most cases of DNA extraction from bones, although it does not provide pure DNA. DNA extraction from aggregates removes inhibitors much better and is also a good method of choice when identity determination of exhumed remains is necessary. In the case of not buried bones (remains found outside) total demineralisation or phenol–chloroform protocols are more efficient for successful DNA extraction.
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