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Preparation of Formalin-fixed Paraffin-embedded Tissue Cores for both RNA and DNA Extraction
Published on: August 21, 2016
A simple method for RNA isolation from formalin-fixed and paraffin-embedded lymphatic tissues
Tajana Körbler1, Marica Grsković, Marija Dominis
1Laboratory of Cellular and Molecular Immunology, Division of Molecular Medicine, Ruder Bosković Institute, Zagreb, Croatia. korbler@rudjer.irb.hr
Insights
Researchers developed a method to isolate mRNA from formalin-fixed, paraffin-embedded lymphatic tissues. This technique enables gene expression analysis in archival samples, crucial for studying lymphoproliferative diseases.
Area of Science:
- Immunology
- Molecular Biology
- Hematology
Background:
- Gene activation is key to lymphoid cell differentiation.
- Analyzing gene expression in lymphoproliferative diseases is challenging due to limited fresh tissue availability.
- Formalin-fixed, paraffin-embedded tissues are common but difficult for molecular analysis.
Purpose of the Study:
- To establish a reliable method for isolating mRNA from archival formalin-fixed, paraffin-embedded lymphatic tissues.
- To enable the study of gene expression, specifically transcription factors, in historical tissue samples.
- To overcome the limitations of fresh tissue scarcity in lymphoproliferative disease research.
Main Methods:
- Adaptation and modification of existing protocols for mRNA isolation.
- Utilized a combination of established methods with a modified phenol/chloroform extraction.
- Applied the developed method to formalin-fixed, paraffin-embedded lymphatic tissue specimens.
Main Results:
- Successfully isolated mRNA from archival formalin-fixed, paraffin-embedded lymphatic tissues.
- Demonstrated the feasibility of screening gene expression analysis using this method.
- Identified potential for studying Ikaros family transcription factors in historical samples.
Conclusions:
- A practical method for mRNA isolation from archival tissues was established.
- This technique facilitates gene expression screening in lymphoproliferative diseases.
- Further research can leverage archival tissues for immunological and hematological studies.
Abstract:
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.
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