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Related Experiment Videos

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
PubMed
Summary
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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:

Related Experiment Videos

  • 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.