title: Deparaffinization with mineral oil: a simple procedure for extraction of high-quality DNA from archival formalin-fixed paraffin-embedded samples.
abstract: Extracting DNA from formalin-fixed paraffin-embedded (FFPE) archival samples remains difficult. Successful polymerase chain reactions (PCR) with DNA extracted from FFPE samples is still very low. We extracted DNA from 12 recent and old archival FFPE bone marrow trephine biopsies by use of a simple protocol on the basis of deparaffinization with molecular biology-grade mineral oil followed by DNA extraction with the Qiagen FFPE kit. Comparison of this deparaffinization method with standard protocols, for example, xylene or Hemo-D with subsequent rehydration using graded ethanols, was investigated. The quality and quantity of extracted DNA were tested by a combination of ultraviolet spectroscopy, analysis on a Caliper LabChip GX, and real-time PCR combined with high-resolution melt analysis. Highest quality PCR-amplifiable DNA was obtained by deparaffinization with mineral oil, whereas more variable results were obtained for the other 2 deparaffinization procedures. This result was confirmed by real-time PCR and high-resolution melt analysis. Besides improvements in the quality of extracted DNA, use of mineral oil for deparaffinization has the added benefit of decreased time (20 vs. 75 min) and a significant reduction of hands-on labor (1 step vs. multiple hands-on centrifugation and decanting steps).
authors.affiliation: *Department of Pathology †Associated Regional University Pathologists (ARUP) Laboratories, University of Utah, Salt Lake City, UT.
authors.forename: Nahla
authors.initials: N
authors.lastname: Heikal
authors.forename: Roberto H
authors.initials: RH
authors.lastname: Nussenzveig
authors.forename: Archana M
authors.initials: AM
authors.lastname: Agarwal
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D017931:DNA Primers; D005557:Formaldehyde; D008899:Mineral Oil / Q000737:chemistry*; D016612:Paraffin Embedding; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D005557:Formaldehyde; D008899:Mineral Oil; D004247:DNA