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

Dekairelle, Anne-France;Van Der Vorst, Sébastien;Tombal, Bertrand;Gala, Jean-Luc
(2007) Clinical Chemistry and Laboratory Medicine : Associated with FESCC and IFCC — Vol. 45, n° 10, p. 1283-1287 (2007)

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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 degrees 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 degrees 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 kappa<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 degrees 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.
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Dekairelle, A.-F., Van Der Vorst, S., Tombal, B., & Gala, J.-L. (2007). Preservation of RNA for functional analysis of separated alleles in yeast: comparison of snap-frozen and RNALater solid tissue storage methods. Clinical Chemistry and Laboratory Medicine : Associated with FESCC and IFCC, 45(10), 1283-1287. https://doi.org/10.1515/CCLM.2007.281 (Original work published 2007)