title: DNA extraction from formalin-fixed material.
abstract: 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.
authors.affiliation: Natural History Museum of Denmark, University of Copenhagen, Østervoldgade 5-7, 1350 Copenhagen, Denmark.
authors.forename: Paula F
authors.initials: PF
authors.lastname: Campos
authors.forename: Thomas M P
authors.initials: TM
authors.lastname: Gilbert
mesh_terms: D000468:Alkalies / Q000737:chemistry; D000818:Animals; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D005404:Fixatives / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry*; D006358:Hot Temperature; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods; D011506:Proteins / Q000737:chemistry; D016707:Tissue Fixation / Q000379:methods
chemical_list: D000468:Alkalies; D003432:Cross-Linking Reagents; D005404:Fixatives; D011506:Proteins; D005557:Formaldehyde; D004247:DNA