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Preservation of RNA for functional analysis of separated alleles in yeast: comparison of snap-frozen and RNALater® solid tissue storage methods
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Anne-France Dekairelle
Veröffentlicht/Copyright:
10. Oktober 2007
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.
Keywords: functional analysis of separated alleles in yeast (FASAY); RNALater®; RNA preservation; snap-frozen
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
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Schlagwörter für diesen Artikel
functional analysis of separated alleles in yeast (FASAY);
RNALater®;
RNA preservation;
snap-frozen
Artikel in diesem Heft
- Atrial natriuretic peptide and related peptides
- The −374A allele of the receptor for advanced glycation end products (RAGE) gene promoter is a protective factor against cardiovascular lesions in type 2 diabetes mellitus patients
- Biochemical properties of endogenous presenilin 1 and presenilin 2 in cultured human B-lymphocytes
- CD36 polymorphism and its relationship with body mass index and coronary artery disease in a Korean population
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- Mother-child double incompatibility at vWA and D5S818 loci in paternity testing
- Matrix metalloprotease-2 and -9 concentration and activity in serum and culture medium samples: a methodological reappraisal
- Are changes in blood-ethanol concentration during storage analytically significant? Importance of method imprecision
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