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Extraction of high-integrity RNA suitable for microarray gene expression analysis from long-term stored human thyroid
Martin A Walter1, Dalma Seboek, Philippe Demougin
1Institute of Nuclear Medicine, University Hospital Basel, Switzerland. m.a.walter@gmx.net
Pathology
|June 7, 2006
Summary
High-integrity RNA was successfully extracted from human thyroid tissues stored for over a decade. This breakthrough enables gene expression profiling in long-term preserved samples, offering new research avenues.
Area of Science:
- Molecular Biology
- Genomics
- Pathology
Background:
- Isolating high-quality RNA from long-term stored thyroid tissues is crucial for gene expression profiling.
- Previously, RNA degradation limited studies on aged tissue samples.
Purpose of the Study:
- To describe the successful extraction of high-integrity RNA from human thyroid tissues stored for over a decade.
- To evaluate the suitability of extracted RNA for gene expression profiling.
Main Methods:
- Processed 79 fresh-frozen thyroid tissue samples (stored 1-16 years) using standard precipitation and column filtration.
- Assessed RNA integrity via electrophoresis (RNA Integrity Number - RIN) and gene expression profiling (GAPDH 3'/5' ratio, Affymetrix present calls).
Main Results:
- Achieved a median RNA yield of 1.9 microg/mg tissue across various thyroid conditions.
- Obtained a median RIN of 8.6, indicating high RNA integrity.
- Demonstrated a median GAPDH 3'/5' ratio of 1.43 and 48.1% present calls, suitable for expression analysis.
Conclusions:
- High-integrity RNA can be reliably extracted from long-term stored fresh-frozen human thyroid tissues.
- The extracted RNA is suitable for gene expression profiling, regardless of tissue storage duration or type.