The tissue bank at the Windber Research Institute focuses on collecting and storing well annotated and high quality tissue and blood samples. In order to demonstrate that the samples are of the highest quality, we do biospecimen research on samples in house to test different processing methods or storage conditions and determine which are best. Pre-analytical variables that could affect downstream research processes that utilize DNA or RNA are studied in ongoing projects at our tissue bank. High DNA/RNA concentration and good yield, integrity and purity are important in all experiments.
One such experiment is our biospecimen research project where we looked at the integrity and purity of RNA that was isolated from breast tissue collected from reductive mammoplasties. Tissue specimens were processed by flash freezing or embedding in optimal cutting temperature (OCT) medium and either immediately stored at -80°C or kept at room temperature over varying time intervals. RNA isolation was performed on all samples and the Nanodrop spectrophotometer was used to determine the concentration and purity of the RNA. RNA integrity number was also determined using the Agilent Bioanaylzer. Since laser capture microdissection is sometimes an integral part of tissue analysis, we also determined its effect on RNA integrity. Quantitative Real-Time PCR was done for comparative measurement of the expression of four house keeping genes: B-actin, glyceraldehyde-3-phosphate dehydrogenase, cyclophilin A and porphobilinogen deaminase. Results of this study can be viewed on the additional poster.
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