RNA quality in fresh frozen prostate tissue from patients operated with radical prostatectomy.
Author(s): Bertilsson H, Angelsen A, Viset T, Anderssen E, Halgunset J
Publication: Scand J Clin Lab Invest, 2010, Vol. 70, Page 45-53
PubMed ID: 20073672 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of blood loss during surgery, sampling location, and automated extraction on RNA yield and integrity from frozen prostatectomy specimens. Slices of prostate specimens were taken immediately after surgery and snap-frozen in liquid nitrogen. 5 mm and 3 mm cores were obtained using an electrical drill for automated and manual analysis, respectively. The elution volumes for the 38 automated and 5 manual specimens were 50 and 100 uL, respectively.
Summary of Findings:
Manual RNA extraction resulted in a higher average RNA yield (29.5 ng/uL versus 24.2 ng/uL) and RNA integrity number (RIN) (8.7 versus 5.5) than automated RNA extraction. RINs were higher in specimens collected from patients with minimal blood loss during surgery than those who lost significant amounts of blood (p<0.02). RINs were also higher in prostate specimens with little stroma compared to those that were 80% or more stroma (p=0.03). There was a trend toward higher RINs in cores from the specimen center compared to cores from the periphery.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform RNA Spectrophotometry RNA Automated electrophoresis/Bioanalyzer Morphology H-and-E microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Analyte isolation method Automated MagNa Pure method
Manual HighPure RNA tissue kit
Biospecimen Aliquots and Components Biospecimen heterogeneity Biospecimen core
Biospecimen periphery
Rich in connective tissue
Rich in epithelial cells
Preaquisition Blood loss amount Minor (<1000 mL)
Significant (>1000 mL)