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Does vacuum centrifugal concentration reduce yield or quality of nucleic acids extracted from FFPE biospecimens?
- Sánchez, Ignacio, Betsou, Fay, Mathieson, William
- Analytical biochemistry 2019 v.566 pp. 16-19
- DNA, RNA, fluorescence emission spectroscopy, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, temperature
- Vacuum centrifugal (SpeedVac) concentration is commonly applied to nucleic acids extracted from formalin-fixed paraffin-embedded (FFPE) sections, but with an unknown impact. We concentrated DNA and RNA from FFPE biospecimens using different time/temperature SpeedVac combinations of up to 30 min concentration at 45 °C, then used spectrophotometry, spectrofluorometry, RIN, PERM, DV200, qRT-PCR, DIN and the Illumina FFPE QC Assay to assess the changes in quantity, purity and integrity induced by the concentration process. We found the effects of SpeedVac concentration to be inconsequential, but an aliquot of elution buffer should be concentrated for use as the blank in spectrophotometry assays.