COMBINED TECHNIQUES OF AUTOMATED IMAGE-GUIDED CORING AND MAGNETIC BEADS BASED PURIFICATION TECHNOLOGY IMPROVES CONCENTRATION OF HIGH SENSITIVITY RNA EXTRACTED FROM FFPE TISSUES
| Autor: | Andrei Cumpanas, Anca Maria Cimpean, Ariana Barmayoun, Alin Adrian Cumpanas, Ovidiu Ferician, Alis Dema, Marius Raica |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: | |
| Zdroj: | Research and Clinical Medicine, Vol I, Iss Suppl.2, Pp 25-25 (2017) |
| ISSN: | 2537-5393 2360-1124 |
| Popis: | Usually, RNA extractions from FFPE specimens have been performed by collecting the tissues based on usually sectioning or by laser capture microdissection followed by RNA extraction manually performed. In most cases the RNA quantity is too low for reverse transcription and real time polymerase chain reaction (RT PCR). Several reasons have been discussed to be responsible for the fail of a proper RNA extraction from FFPE specimens, from the tisular damage caused by formalin to the nuclear integrity or the age of the paraffin blocks. MATERIAL AND METHODS. We performed a fully automated workflow of RNA extraction, based on automated tissue microarray (TMA) specimens collection followed by magnetic beads based purification technology on 36 randomly choosed archival specimens of kidney cancer. The unique variable of the procedure was the core numbers. RESULTS. From all specimens of 4 cores group RNA was succesfully extracted, its concentration ranging between 1350 to 9420 ng/ml. Eight out of 24 specimens of 2 cores groups (30%) showed detectable levels of RNA (ranged between 460 to 3420 ng/ml), 70 % of them being out of standard range. There is a statistically significant difference (P-value = 0,00002) between the number of cores (4 versus 2) and RNA concentration. CONCLUSIONS. Age of the specimens or formalin do not influence High Sensitivity RNA concentration extracted from FFPE specimens. TMA image-guided coring preserves nuclear integrity and thus may improve RNA extraction. RNA concentration may be also improved by association of the automated image guided coring with semi automated magnetic beads based techniques and by adjusting core numbers. |
| Databáze: | OpenAIRE |
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