Accessible Unlicensed Requires Authentication Published by De Gruyter October 10, 2007

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

Anne-France Dekairelle, Sébastien Van der Vorst, Bertrand Tombal and Jean-Luc Gala
From the journal

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.


Corresponding author: Jean Luc Gala, MD, PhD, Centre for Applied Molecular Technology, BP 30.46, 30 Clos Chapelle aux Champs, 1200 Bruxelles, Belgium Phone: +32-2-764-3165, Fax: +32-2-764-3166,

Received: 2007-4-12
Accepted: 2007-6-1
Published Online: 2007-10-10
Published in Print: 2007-10-01

©2007 by Walter de Gruyter Berlin New York