Comparison of methods in the recovery of nucleic acids from archival formalin-fixed paraffin-embedded autopsy tissues

被引:88
|
作者
Okello, John B. A. [1 ]
Zurek, Jaymi [1 ]
Devault, Alison M. [1 ]
Kuch, Melanie [1 ]
Okwi, Andrew L. [2 ]
Sewankambo, Nelson K. [3 ]
Bimenya, Gabriel S. [2 ]
Poinar, Debi [1 ]
Poinar, Hendrik N. [1 ,4 ,5 ]
机构
[1] McMaster Univ, Dept Anthropol, McMaster Ancient DNA Ctr, Hamilton, ON L8S 4L9, Canada
[2] Makerere Univ, Coll Hlth Sci, Sch Biomed Sci, Dept Pathol, Kampala, Uganda
[3] Makerere Univ, Coll Hlth Sci, Sch Med, Clin Epidemiol Unit, Kampala, Uganda
[4] McMaster Univ, Dept Pathol & Mol Med, Hamilton, ON L8N 3Z5, Canada
[5] McMaster Univ, Michael DeGroote Inst Infect Dis Res, Hamilton, ON L8N 3Z5, Canada
基金
加拿大健康研究院;
关键词
Archival FFPE tissues; DNA/RNA extraction; Histopathology; Inhibition; Nucleic acid; Quantitative PCR; DNA EXTRACTION METHODS; GENE-EXPRESSION; RNA EXTRACTION; PCR PRODUCTS; FIXATION; AMPLIFICATION; OPTIMIZATION; DEGRADATION; TIME; PURIFICATION;
D O I
10.1016/j.ab.2010.01.014
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Archival formalin-fixed paraffin-embedded (FFPE) human tissue collections are typically in poor states of storage across the developing world. With advances in biomolecular techniques, these extraordinary and virtually untapped resources have become an essential part of retrospective epidemiological studies. To successfully use such tissues in genomic studies, scientists require high nucleic acid yields and purity. In spite of the increasing number of FFPE tissue kits available, few studies have analyzed their applicability in recovering high-quality nucleic acids from archived human autopsy samples. Here we provide a study involving 10 major extraction methods used to isolate total nucleic acid from FFPE tissues ranging in age from 3 to 13 years. Although all 10 methods recovered quantifiable amounts of DNA, only 6 recovered quantifiable RNA, varying considerably and generally yielding lower DNA concentrations. Overall, we show quantitatively that TrimGen's WaxFree method and our in-house phenol-chloroform extraction method recovered the highest yields of amplifiable DNA, with considerable polymerase chain reaction (PCR) inhibition, whereas Ambion's RecoverAll method recovered the most amplifiable RNA. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码:110 / 117
页数:8
相关论文
共 50 条
  • [1] Comparison of Methods for the Extraction of DNA from Formalin-Fixed, Paraffin-Embedded Archival Tissues
    Senguven, Burcu
    Baris, Emre
    Oygur, Tulin
    Berktas, Mehmet
    INTERNATIONAL JOURNAL OF MEDICAL SCIENCES, 2014, 11 (05): : 494 - 499
  • [2] Comparison of nucleic acid extraction methods for formalin-fixed paraffin-embedded tissues
    Rich, R. G.
    Hafez, M. J.
    Gocke, C.
    Murphy, K. M.
    JOURNAL OF MOLECULAR DIAGNOSTICS, 2007, 9 (05): : 701 - 701
  • [3] Recovery of proteins from formalin-fixed paraffin-embedded tissues
    O'Leary, Timothy J.
    Fowler, Carol
    Fabris, Dan
    Mason, Jeffrey T.
    BIOPHYSICAL JOURNAL, 2007, : 388A - 388A
  • [4] Analysis of amelogenin by PCR in the DNA extracted from archival formalin-fixed, paraffin-embedded autopsy tissues
    Funabashi, K. S.
    Barcelos, D.
    Visona, I.
    Silva, M. S.
    Godoy, C. D.
    Franco, M. F.
    Iwamura, E. S. M.
    HISTOPATHOLOGY, 2010, 57 : 197 - 197
  • [5] Extraction of DNA from formalin-fixed paraffin-embedded tissues: A comparison of methods
    Carter, A. B.
    Melan, M. A.
    Sheikh, H.
    Kant, J. A.
    JOURNAL OF MOLECULAR DIAGNOSTICS, 2007, 9 (05): : 702 - 702
  • [6] Comparison of DNA extraction methods of formalin fixed, paraffin-embedded archival tissues
    Senguven, B.
    Baris, E.
    Oygur, T.
    VIRCHOWS ARCHIV, 2012, 461 : S171 - S171
  • [7] 'Tissue surrogates' as a model for archival formalin-fixed paraffin-embedded tissues
    Fowler, Carol B.
    Cunningham, Robert E.
    O'Leary, Timothy J.
    Mason, Jeffrey T.
    LABORATORY INVESTIGATION, 2007, 87 (08) : 836 - 846
  • [8] The quality of DNA isolated from autopsy formalin-fixed and formalin-fixed paraffin-embedded tissues: study of 1662 samples
    Katarina Vitošević
    Danijela Todorović
    Živana Slović
    Tatjana Varljen
    Ivana Radaković
    Dušan Radojević
    Vanja Čanović
    Miloš Todorović
    Molecular Biology Reports, 2023, 50 : 6323 - 6336
  • [9] Spatially exploring RNA biology in archival formalin-fixed paraffin-embedded tissues
    Bai, Zhiliang
    Zhang, Dingyao
    Gao, Yan
    Tao, Bo
    Zhang, Daiwei
    Bao, Shuozhen
    Enninful, Archibald
    Wang, Yadong
    Li, Haikuo
    Su, Graham
    Tian, Xiaolong
    Zhang, Ningning
    Xiao, Yang
    Liu, Yang
    Gerstein, Mark
    Li, Mingyao
    Xing, Yi
    Lu, Jun
    Xu, Mina L.
    Fan, Rong
    CELL, 2024, 187 (23)
  • [10] The quality of DNA isolated from autopsy formalin-fixed and formalin-fixed paraffin-embedded tissues: study of 1662 samples
    Vitosevic, Katarina
    Todorovic, Danijela
    Slovic, Zivana
    Varljen, Tatjana
    Radakovic, Ivana
    Radojevic, Dusan
    Canovic, Vanja
    Todorovic, Milos
    MOLECULAR BIOLOGY REPORTS, 2023, 50 (08) : 6323 - 6336