Comparison of DNA and RNA extraction methods for mummified tissues.
Nucleic acids extracted from mummified tissues are valuable materials for the study of ancient human beings. Significant difficulty in extracting nucleic acids from mummified tissues has been reported due to chemical modification and degradation. The goal of this study was to determine a method that is more efficient for DNA and RNA extraction from mummified tissues. Twelve mummy specimens were analyzed with 9 different nucleic acid extraction methods, including guanidium thiocyanate (GTC) and proteinase K/detergent based methods prepared in our laboratory or purchased. Glyceraldehyde 3-phosphate dehydrogenase DNA and beta-actin RNA were used as markers for the presence of adequate DNA and RNA, respectively, for PCR and RT-PCR amplification. Our results show that 5 M GTC is more efficient of releasing nucleic acids from mummified tissue than proteinase K/detergent, and phenol/chloroform extraction with an additional chloroform step is more efficient than phenol/chloroform along. We were able to isolate DNAs from all 12 specimens and RNAs from 8 of 12 specimens, and the nucleic acids were sufficient for PCR and RT-PCR analysis. We further tested hepatitis viruses including hepatitis B virus, hepatitis C virus, hepatitis G virus, and TT virus DNA, and fail to detect these viruses in all 12 specimens.
Konomi
Nami
N
Department of Pathology and Dermatology, Mount Sinai School of Medicine, New York University, New York, NY 10029, USA.
Lebwohl
Eve
E
Zhang
David
D
D002725:Chloroform; D004247:DNA / Q000302:isolation & purification*; D019286:Endopeptidase K; D005260:Female; D006146:Guanidines; D006518:Hepatitis Viruses / Q000302:isolation & purification; D049690:History, Ancient; D049691:History, Medieval; D006801:Humans; D008297:Male; D009106:Mummies / Q000473:pathology* / Q000821:virology; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification*; D013861:Thiocyanates; D022783:Torque teno virus / Q000302:isolation & purification
D006146:Guanidines; D013861:Thiocyanates; C054436:guanidine thiocyanate; D012313:RNA; D002725:Chloroform; D004247:DNA; D019286:Endopeptidase K