Celoten zapis

BS-ID zapisa
156603
COBISS-ID
28186585
Tip dokumenta
primarni članek
Avtorji
Zupanič-Pajnič I.
Naslov
Visoko učinkovita metoda ekstrakcije DNA iz skeletnih ostankov
Vir
Zdrav Vestn 2011; 80(3):171-81.
Vsebinske oznake
human
Deskriptorji (glavni)
FORENSIC ANTHROPOLOGY
BONE AND BONES
TOOTH
DNA
DNA FINGERPRINTING
Deskriptorji (stranski)
POLYMERASE CHAIN REACTION
DNA, MITOCHONDRIAL
Abstrakt
Background: This paper precisely describes the method of DNA extraction developed to acquire high quality DNA from the Second World War skeletal remains. The same method is also used for molecular genetic identification of unknown decomposed bodies in routine forensic casework where only bones and teeth are suitable for DNA typing. We analysed 109 bones and two teeth from WWII mass graves in Slovenia. Methods: We cleaned the bones and teeth, removed surface contaminants and ground the bones into powder, using liquid nitrogen. Prior to isolating the DNA in parallel using the BioRobot EZ1 (Qiagen), the powder was decalcified for three days. The nuclear DNA of the samples were quantified by real-time PCR method. We acquired autosomal genetic profiles and Y-chromosome haplotypes of the bones and teeth with PCR amplification of microsatellites, and mtDNA haplotypes 99. For the purpose of traceability in the event of contamination, we prepared elimination data bases including genetic profiles of the nuclear and mtDNA of all persons who have been in touch with the skeletal remains in any way. Results: We extracted up to 55 ng DNA/g of the teeth, up to 100 ng DNA/g of the femurs, up to 30 ng DNA/g of the tibias and up to 0.5 ng DNA/g of the humerus. The typing of autosomal and YSTR loci was successful in all of the teeth, in 98 % dekalof the femurs, and in 75 % to 81 % of the tibias and humerus. The typing of mtDNA was successful in all of the teeth, and in 96 % to 98 % of the bones. Conclusions: We managed to obtain nuclear DNA for successful STR typing from skeletal remains that were over 60 years old . The method of DNA extraction described here has proved to be highly efficient. We obtained 0.8 to 100 ng DNA/g of teeth or bones and complete genetic profiles of autosomal DNA, Y-STR haplotypes, and mtDNA haplotypes from only 0.5g bone and teeth samples.
URL
http://szd.si/user_files/vsebina/Zdravniski_Vestnik/2011/marec/171-81.pdf