A simple method for RNA isolation from formalin-fixed and paraffin-embedded lymphatic tissues.
Gene activation that lies beneath lymphoid cell differentiation has been one of the most explored issues in immunology in the recent years. However, the analysis of this molecular event in lymphoproliferative diseases is often hampered by the lack of fresh material. Most tissues available for routine histological investigation are formalin fixed and paraffin embedded. Gene expression in such specimens could be analyzed using reverse transcription of mRNA and the polymerase chain reaction (RT-PCR). Therefore we adjusted and established a method for mRNA isolation from such specimens by a combination of previously reported protocols and a modification of the phenol/chloroform extraction method. Given the significance of transcription factors in the human hemopoietic system, we investigated whether mRNA could be successfully isolated from archival tissue for a study on expression of Ikaros family transcription factors in lymphatic tissue. Although quantitative analysis of RNA isolated from archival tissue is probably not feasible due to the unpredictable degree of RNA isolation varying from sample to sample, we show here that screening analysis is possible and simple.
Körbler
Tajana
T
Laboratory of Cellular and Molecular Immunology, Division of Molecular Medicine, Ruder Bosković Institute, Zagreb, Croatia. korbler@rudjer.irb.hr
Grsković
Marica
M
Dominis
Marija
M
Antica
Mariastefania
M
D017931:DNA Primers / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005404:Fixatives; D005557:Formaldehyde; D006689:Hodgkin Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008198:Lymph Nodes / Q000737:chemistry* / Q000473:pathology; D008228:Lymphoma, Non-Hodgkin / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D016612:Paraffin Embedding; D012333:RNA, Messenger / Q000378:metabolism; D012334:RNA, Neoplasm / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014157:Transcription Factors
D017931:DNA Primers; D005404:Fixatives; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D014157:Transcription Factors; D005557:Formaldehyde