title: Comparison of protocols for DNA extraction from long-term preserved formalin fixed tissues.
abstract: The current study compared the applicability of protocols to extract DNA from formalin fixed heart tissues that have been preserved for more than 50 years. Ten methods were tested: a cetyltrimethylammonium bromide (CTAB) standard protocol, seven variants of this standard protocol, and two commercial kits. In the case of younger specimens (fixed in 1951, 1934, or 1914), extracts with DNA concentrations ≥ 10.0 ng/μl were obtained with the standard CTAB protocol, two variants of the standard protocol including prolonged tissue digestion (72 h instead of 1-2h), and a commercial kit particularly recommended for DNA extraction from formalin fixed paraffin embedded tissues (FFPE Kit). With the FFPE Kit, DNA could also be extracted from older tissues (fixed in 1893, 1850/1851, or before 1820). In general, the purity of the DNA extracts, assessed from the ratio of the absorbance at 260 and 280 nm, was not very high. In spite of their rather low purity, the DNA extracts could, however, be used to amplify a 122-bp sequence and, in most cases, also a 171-bp sequence of the gene coding for human albumin by the polymerase chain reaction (PCR).
authors.affiliation: Department of Analytical Chemistry, University of Vienna, 1090 Vienna, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Paireder
authors.forename: Bettina
authors.initials: B
authors.lastname: Werner
authors.forename: Josef
authors.initials: J
authors.lastname: Bailer
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Werther
authors.forename: Erich
authors.initials: E
authors.lastname: Schmid
authors.forename: Beatrix
authors.initials: B
authors.lastname: Patzak
authors.forename: Margit
authors.initials: M
authors.lastname: Cichna-Markl
mesh_terms: D000818:Animals; D005591:Chemical Fractionation / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D005557:Formaldehyde / Q000737:chemistry*; D006801:Humans; D009206:Myocardium / Q000737:chemistry*; D013552:Swine; D013997:Time Factors; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D005557:Formaldehyde; D004247:DNA
keywords: Archival museum specimens; DNA extraction; Formalin fixed tissue; Heart; PCR amplifiability