Comparison of DNA and RNA extraction methods for mummified tissues

被引:29
|
作者
Konomi, N
Lebwohl, E
Zhang, D
机构
[1] NYU, Mt Sinai Sch Med, Mol Pathol Lab, Dept Pathol, New York, NY 10021 USA
[2] NYU, Mt Sinai Sch Med, Dept Dermatol, New York, NY 10021 USA
关键词
PCR; RT-PCR; hepatitis; guanidium thiocyanide;
D O I
10.1006/mcpr.2002.0441
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 P-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 5M 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. (C) 2002 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:445 / 451
页数:7
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