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
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