Preservation of RNA for functional analysis of separated alleles in yeast: comparison of snap-frozen and RNALater® solid tissue storage methods

Anne-France Dekairelle 1 , 1 , Sébastien Van der Vorst 2 , 2 , Bertrand Tombal 3 , 3  and Jean-Luc Gala 4 , 4
  • 1 Centre for Applied Molecular Technology, Université catholique de Louvain, Brussels, Belgium
  • 2 Centre for Applied Molecular Technology, Université catholique de Louvain, Brussels, Belgium The first two authors contributed equally to this work.
  • 3 Service d'Urologie, Cliniques universitaires Saint-Luc, Brussels, Belgium
  • 4 Centre for Applied Molecular Technology, Université catholique de Louvain, Brussels, Belgium and Defence Laboratories Department, Belgian Armed Forces, Brussels, Belgium

Abstract

Background: The aim of the present study was to compare RNALater® with the usual method of liquid nitrogen snap freezing as a surrogate mRNA preservation method for functional analysis of separated alleles in yeast (FASAY).

Methods: A total of 81 patients with transitional cell carcinoma of the bladder underwent fresh tissue biopsies directly transferred into RNALater® and stored at room temperature or at 4°C for increasing time intervals until RNA processing. From this cohort of patients, 53 paired snap-frozen and RNALater® preservative-suspended tissues were obtained. Samples immediately frozen in liquid nitrogen were further stored at −80°C.

Results: Of the 81 RNALater® samples, 14 were not processed for FASAY because of RNA degradation. Of the remaining 67 samples, 15 (22%) were FASAY-positive. Identical FASAY results were found for 50 of 53 (94.4%) paired samples and the percentage of red yeast colonies was highly correlated (Cohen's κ<0.82; p<0.00001). A single p53 missense mutation was found in each of the three discordant positive FASAY and was identical in each concordant positive sample (10/53). Storing samples in RNALater® at room temperature for 3 days and at 4°C for less than 1 month provided high-quality mRNA suitable for FASAY.

Conclusions: Our results demonstrate that RNALater® is a suitable and flexible alternative to snap freezing for FASAY analysis.

Clin Chem Lab Med 2007;45:1283–7.

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Clinical Chemistry and Laboratory Medicine ( CCLM) publishes articles on novel teaching and training methods applicable to laboratory medicine. CCLM welcomes contributions on the progress in fundamental and applied research and cutting-edge clinical laboratory medicine. It is one of the leading journals in the field, with an impact factor of over three. CCLM is the official journal of nine national clinical societies and associated with EFLM.

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