DNA extraction from formalin-fixed material

The principal challenges facing PCR-based analyses of DNA extracted from formalin-fixed materials are fragmentation of the DNA and cross-linked protein-DNA complexes. Here, we present an efficient protocol to extract DNA from formalin-fixed or paraffin-embedded tissues (FFPE). In this protocol, prot...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 840; p. 81
Main Authors Campos, Paula F, Gilbert, Thomas M P
Format Journal Article
LanguageEnglish
Published United States 01.01.2012
Subjects
Alkalies - chemistry
Animals
Cross-Linking Reagents - chemistry
DNA - chemistry
DNA - isolation & purification
Fixatives - chemistry
Formaldehyde - chemistry
Hot Temperature
Paraffin Embedding
Polymerase Chain Reaction - methods
Proteins - chemistry
Tissue Fixation - methods
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Summary:The principal challenges facing PCR-based analyses of DNA extracted from formalin-fixed materials are fragmentation of the DNA and cross-linked protein-DNA complexes. Here, we present an efficient protocol to extract DNA from formalin-fixed or paraffin-embedded tissues (FFPE). In this protocol, protein-DNA cross-links are reversed using heat and alkali treatment, yielding significantly longer fragments and larger amounts of PCR-amplifiable DNA than standard DNA extraction protocols.
ISSN:1940-6029
DOI:10.1007/978-1-61779-516-9_11