engleski

Isolation of RNA from archive paraffin-embedded lymphatic tissue specimens

The analysis of lymphoproliferative diseases by molecular methods is often hampered by the lack of fresh material. Most tissues available for routine pathological investigation, as well as comprehensive studies are formalin-fixed and paraffin-embedded. Gene expression in such specimens could be analysed using reverse transcription of RNA and polymerase chain reaction (RT-PCR). This method requires, above all, high quality RNA isolation. In this study, we isolated RNA from 20 formalin-fixed and paraffin-embedded lymph node tissue samples by modifying the phenol/chloroform extraction method. We assessed the quality of isolated RNA by agarose-gel electrophoresis and compared it to the RNA isolated from the fresh material. Our results show that ribosomal RNA from archive material is highly degraded comparing to RNA from fresh material. However, mRNA seems to be intact since we managed to amplify a control house keeping b-actin gene by means of RT-PCR. Therefore, in the light of gene expression investigation, our method could be successfully applied for extraction of RNA from formalin-fixed and paraffin-embedded specimens.

RNA isolation; paraffin-embedded; lymphatic tissue

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