Molecular analysis of a collection of clinical specimens stored at 4 degrees C as an alternative to snap-freezing.

Author(s): Sewart S, Barraclough R, Rudland PS, West CR, Barraclough DL

Publication: Int J Oncol, 2009, Vol. 35, Page 381-6

PubMed ID: 19578753 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study investigated the effects of storing freshly procured surgical specimens at 4 degrees C for up to 24 h prior to preservation. DNA, RNA, and protein indices were compared among 36 case-matched specimens snap frozen in liquid nitrogen within 30 min of surgery and stored at -140 degrees C until analysis.

   Summary of Findings:

   PCR and protein banding patterns were identical among specimens stored at 4 degrees C for up to 24 h and those that were snap frozen in liquid nitrogen. RNA yields did not differ significantly among differentially preserved case-matched specimens, nor did the results of Northern blot analysis for which intact 1.5 kb transcripts and similar degrees of degradation were observed among differentially preserved specimens. Transcript-specific expression, determined by RT-PCR and cDNA microarray, were not affected by specimen storage at 4 degrees C for up to 24 h compared to case-matched snap frozen controls, indicating that expression of 190 mRNA transcripts assessed were not affected by storage of fresh cancerous specimens at 4 degrees C for up to 24 h.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Other Preservative
   • Frozen

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   DNA	PCR
   RNA	Spectrophotometry
   RNA	Northern blot
   RNA	RT-PCR
   RNA	DNA microarray
   Protein	1D/2D gels

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Refrigeration Snap frozen
   Biospecimen Acquisition	Cold ischemia time	30 min 18-24 h
   PCR Specific	Targeted nucleic acid	S100A4 gene TATA box
   RT-PCR Specific	Targeted nucleic acid	HPRT
   RT-PCR Specific	Length of gene fragment	167 bp 246 bp 367 bp 473 bp 656 bp

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