Comparison of the DNA Preservation in Neutral-Buffered Formalin Fixed Paraffin-Embedded Tissue and in Non-Buffered Formalin Fixed Paraffin-Embedded Tissue
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Background: The preservation of optimized DNA and its extraction from formalin-fixed, paraffin-embedded (FFPE) tissues are important issues. There has been some doubt over whether 10% neutral-buffered formalin is an ideal fixation solution for DNA preservation over non-buffered for¬malin, as conventionally recommended. In this study, the correlation between the efficiency of DNA extraction from FFPE tissues and buffered formalin was evaluated. Methods: Several tis-sues with same conditions except fixatives were fixed in four different formalin solution groups and were routinely processed as paraffin-embedding protocols. DNAs were extracted from four different FFPE tissues that were stored for over 3 months and over 9 months. The quantity and quality of the DNAs were assessed with a NanoDrop ND-1000 spectrophotometer, and the poly-merase chain reaction (PCR) amplification and degradation were analyzed via microchip electro-phoresis. KRAS mutation analysis and microsatellite instability (BAT25) PCR were performed with each sample. Results: The results showed no remarkable difference in the four groups. Conclu-sions: The study findings demonstrate that DNA preservation is fairly unaffected by a neutral buf-fer where there is short formalin manufacture period and an adequate formalin fixation time be-fore embedding in paraffin.
Abstract
Background: The preservation of optimized DNA and its extraction from formalin-fixed, paraffin-embedded (FFPE) tissues are important issues. There has been some doubt over whether 10% neutral-buffered formalin is an ideal fixation solution for DNA preservation over non-buffered for¬malin, as conventionally recommended. In this study, the correlation between the efficiency of DNA extraction from FFPE tissues and buffered formalin was evaluated. Methods: Several tis-sues with same conditions except fixatives were fixed in four different formalin solution groups and were routinely processed as paraffin-embedding protocols. DNAs were extracted from four different FFPE tissues that were stored for over 3 months and over 9 months. The quantity and quality of the DNAs were assessed with a NanoDrop ND-1000 spectrophotometer, and the poly-merase chain reaction (PCR) amplification and degradation were analyzed via microchip electro-phoresis. KRAS mutation analysis and microsatellite instability (BAT25) PCR were performed with each sample. Results: The results showed no remarkable difference in the four groups. Conclu-sions: The study findings demonstrate that DNA preservation is fairly unaffected by a neutral buf-fer where there is short formalin manufacture period and an adequate formalin fixation time be-fore embedding in paraffin.
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