| The isolation of nucleic acids from fixed, paraffin-embedded tissues-which methods are useful when? | |
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MedLine Citation:
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PMID: 17579711 Owner: NLM Status: PubMed-not-MEDLINE |
Abstract/OtherAbstract:
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Museums and pathology collections around the world represent an archive of genetic material to study populations and diseases. For preservation purposes, a large portion of these collections has been fixed in formalin-containing solutions, a treatment that results in cross-linking of biomolecules. Cross-linking not only complicates isolation of nucleic acid but also introduces polymerase "blocks" during PCR. A wide variety of methods exists for the recovery of DNA and RNA from archival tissues, and although a number of previous studies have qualitatively compared the relative merits of the different techniques, very few have undertaken wide scale quantitative comparisons. To help address this issue, we have undertaken a study that investigates the quality of nucleic acids recovered from a test panel of fixed specimens that have been manipulated following a number of the published protocols. These include methods of pre-treating the samples prior to extraction, extraction and nucleic acid purification methods themselves, and a post-extraction enzymatic repair technique. We find that although many of the published methods have distinct positive effects on some characteristics of the nucleic acids, the benefits often come at a cost. In addition, a number of the previously published techniques appear to have no effect at all. Our findings recommend that the extraction methodology adopted should be chosen carefully. Here we provide a quick reference table that can be used to determine appropriate protocols for particular aims. |
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Authors:
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M Thomas P Gilbert; Tamara Haselkorn; Michael Bunce; Juan J Sanchez; Sebastian B Lucas; Laurence D Jewell; Eric Van Marck; Michael Worobey |
Publication Detail:
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Type: Journal Article Date: 2007-06-20 |
Journal Detail:
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Title: PloS one Volume: 2 ISSN: 1932-6203 ISO Abbreviation: PLoS ONE Publication Date: 2007 |
Date Detail:
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Created Date: 2007-06-20 Completed Date: 2007-07-31 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 101285081 Medline TA: PLoS One Country: United States |
Other Details:
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Languages: eng Pagination: e537 Citation Subset: - |
Affiliation:
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Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, Arizona, United States of America. mtpgilbert@gmail.com |
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