| dc.description.abstract | As the genetic basis and gene expression components are being discovered for an increasing
number of diseases and disorders there is a rising interest in using multiomic approaches to
better understand these conditions. The large tissue banks of formalin fixed (FF) and
formalin-fixed paraffin-embedded (FFPE) tissues present a unique opportunity to study rare
conditions and difficult to sample organs such as brains. Current molecular studies on FFPE
look at single molecule extractions, which provide only a partial picture of disease
mechanisms. This study compared the efficacy of TRIzol®, a tri-molecule extraction
method, to that of the traditional single molecule methods for DNA and RNA extraction in
frozen, FF, and FFPE brain tissue using animal models. Optimization of this would allow for
DNA and RNA to be obtained from the same sample providing a more accurate and
comprehensive picture of the mechanism of disease or disorder. The quantity and purity of
the DNA and RNA extracted from frozen, FF, and FFPE brain samples (aged for 3 years)
were determined by spectrophotometry. Quality was determined by real-time PCR (qPCR)
and reverse transcriptase qPCR (RT-qPCR) using beta-2-microglobulin (98bp) and
glyceraldehyde 3-phosphate dehydrogenase (227bp) targets. Our study found no significant
difference in efficacy between TRIzol® and the traditional methods employed in the
literature, indicating that TRIzol® is comparable to the currently used, single molecule
extractions when applied to FF and FFPE brain tissue. This finding requires further
investigation with a larger set of samples, but if supported may help maximize the amount of
information obtained from rare samples as multiple data sets would be obtained from a single
sample. Unexpectedly, this study also observed a higher rate of success in extracting high
quality RNA compared to DNA regardless of extraction method. Due to the low number of
samples, the statistical significance of this observation could not be established however we
feel that it warrants further investigation. | en_US |
