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 in:	Methods in molecular biology (Clifton, N.J.) Vol. 840; p. 81
Main Authors:	Campos, Paula F, Gilbert, Thomas M P
Format:	Journal Article
Language:	English
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