Original Articles
Suphasarang Sirirattanakul1, T
Abstract
Formalin-fixed paraffin embedded (FFPE) tissue is a tissue obtained from dissection of the patients’ lesion and is well preserved as a paraffin block. This specimen is regarded as a wealthy resource for RNA studies in various diseases for the advantages of not only diagnosis and prognosis, but also treatment. The aim of our study was to demonstrate that amplifiable RNA could be recovered from FFPE tissues using paramagnetic bead-based isolation technique. RNA isolated on manual and automated platforms was compared for their integrity, yield and purity. Expression of three housekeeping genes from the isolated RNA products was used as a tool to confirm amplificability of the isolated RNA of different amplicon sizes. The results showed that RNA could be successfully recovered from a 10 μm FFPE tissues of 20 samples. The manual platform gave better result for A260/A280 absorbance ratio (1.93±0.009 vs 1.82 ±0.007; p<0.05), whereas the automated platform showed better result with regard to quantity (19.31±2.530 vs 8.87 μg ±1.468 μg). Using reverse transcription polymerase chain reaction (RTPCR), isolated RNA obtained from both platforms could be achieved with amplicon size up to 656 bp. In this study, we successfully isolated RNA from FFPE tissues using manual and automated platforms under the paramagnetic bead-based scheme with good quality and quantity of the RNA products. Noteworthy, the results also showed longer length of amplified product derived from isolated RNA of FFPE than any other previous reports.