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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Comparison of methods in the recovery of nucleic acids from archival formalin-fixed paraffin-embedded autopsy tissues
Analytical Biochemistry, Volume 400, No. 1, Year 2010
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Description
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. © 2010 Elsevier Inc. All rights reserved.
Authors & Co-Authors
Okello, John Bosco A.
Canada, Hamilton
Mcmaster Ancient Dna Centre
Zurek, Jaymi
Canada, Hamilton
Mcmaster Ancient Dna Centre
Devault, Alison M.
Canada, Hamilton
Mcmaster Ancient Dna Centre
Kuch, Melanie
Canada, Hamilton
Mcmaster Ancient Dna Centre
Okwi, Andrew Livex
Uganda, Kampala
College of Health Sciences
Sewankambo, Nelson K.
Uganda, Kampala
College of Health Sciences
Bimenya, Gabriel S.
Uganda, Kampala
College of Health Sciences
Poinar, Debi
Canada, Hamilton
Mcmaster Ancient Dna Centre
Poinar, Hendrik N.
Canada, Hamilton
Mcmaster Ancient Dna Centre
Canada, Hamilton
Mcmaster University
Canada, Hamilton
Michael G. Degroote Institute for Infectious Disease Research
Statistics
Citations: 123
Authors: 9
Affiliations: 4
Identifiers
Doi:
10.1016/j.ab.2010.01.014
ISSN:
00032697
e-ISSN:
10960309
Research Areas
Genetics And Genomics
Study Design
Cohort Study