Preservation of RNA for Functional Genomic Studies: A Multidisciplinary Tumor Bank Protocol
Author(s): Florell Scott R, Coffin Cheryl M, Holden Joseph A, Zimmermann James W, Gerwells John W, Summers Bradley K, Jones David A, Leachman Sancy A
Publication: Mod Pathol, 2001, Vol. 14, Page 116
PubMed ID: 11235903 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
Comparative histological analysis of tissue samples fixed with RNAlater and other fixation methods (formalin, OCT).
Summary of Findings:
Histological staining, assessed by three pathologists in a blind study, was indistinguishable from other fixation methods (formalin, OCT) in tissue samples preserved in RNAlater for 24 h up to 3 weeks. Note: experimental samples included surgically resected normal skin tissue, as well as several tissue types collected during autopsy (brain, thyroid, lung, heart, skeletal muscle, stomach, small and large intestine, tongue).
Biospecimens
Preservative Types
- RNAlater
- OCT
- Formalin
Diagnoses:
- Normal
- Autopsy
Platform:
Analyte Technology Platform Morphology H-and-E microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
OCT
RNAlater
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Study Purpose
Comparative analysis of immunohistochemical staining of tissue fixed with RNAlater and other fixation methods (formalin, OCT).
Summary of Findings:
Immunohistochemical staining was indistinguishable from other fixation methods (formalin, OCT, TRIzol reagent) in tissue samples preserved in RNAlater for 24 h and stained for CEA, cytokeratin AE 1/3, and vimentin. S-100 staining was not observed in RNAlater fixed epidermal melanocytes or Langerhan's cells, although these cell types were stained in fixative controls. Tissues stored in TRIzol reagent for 24 h exhibited extensive cytolysis and lacked nuclei. Note: experimental samples included surgically resected normal skin tissue, as well as several tissue types collected during autopsy (brain, thyroid, lung, heart, skeletal muscle, stomach, small and large intestine, tongue).
Biospecimens
Preservative Types
- RNAlater
- OCT
- Formalin
Diagnoses:
- Autopsy
- Normal
Platform:
Analyte Technology Platform Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
OCT
RNAlater
Biospecimen Preservation RNA stabilization method RNAlater
Trizol reagent
Immunohistochemistry Specific Targeted peptide/protein CEA
Vimentin
S-100
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Study Purpose
To evaluate RNA integrity in normal skin samples preserved in RNAlater at ambient temperature for 24 h, followed by 4 degrees C for 2-6 weeks in comparison to fresh frozen controls.
Summary of Findings:
RNA extracted from skin samples fixed in RNAlater were superiorly preserved compared to fresh frozen controls, displaying distinct ribosomal bands and high molecular weight transcripts with limited degradation.
Biospecimens
Preservative Types
- RNAlater
- Frozen
Diagnoses:
- Normal
Platform:
Analyte Technology Platform RNA Electrophoresis Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation RNAlater
Snap frozen
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Study Purpose
To determine whether RNA extracted from tissues fixed in RNAlater was intact and sufficient for microarray analysis.
Summary of Findings:
RNA extracted from skin (basal cell carcinoma, squamous cell carcinoma) preserved in RNAlater was successful used in microarray analysis. Northern blot analysis confirmed microarray results, identifying an upregulated 2.5 kb MIG transcript.
Biospecimens
Preservative Types
- RNAlater
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform RNA DNA microarray RNA Northern blot Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Northern blot Specific Targeted nucleic acid MIG