Biological stability of RNA isolated from RNAlater-treated brain tumor and neuroblastoma xenografts

Abstract

Background Advances in molecular biological research have led to identification of prognostic factors such as Trk mRNA expression in primitive neuroectodermal tumors of the CNS and neuroblastoma. To study prospectively the importance of these prognostic factors in large groups of homogeneously treated patients, tumor specimens of good quality must be acquired, preserved, and stored at multiple institutions. Immediate freezing of tumor biopsy samples in liquid nitrogen and storage at −70°C are the most commonly used method of tissue preservation for future RNA analysis. Procedure To evaluate alternative methods of preserving tissue samples for subsequent RNA analysis, we tested a new RNA stabilization solution. Using tumor tissue of two CNS tumor and one neuroblastoma human xenografts, we compared total RNA isolated from tumor tissue stored for 7 days at room temperature in stabilization solution to that of snap‐frozen tissue. The quality of the RNA was studied by spectrophotometry, gel electrophoresis, RT‐PCR, and gene expression profiling. Results No major differences were observed in the quality of RNA isolated from tumor samples stored at room temperature in the RNA stabilization solution compared to snap‐frozen tumor samples stored at −70°C. Conclusions High‐quality RNA can be prepared from tumor tissue stored at room temperature. Whenever snap freezing is not feasible, pieces of tumor tissue can be treated with RNAlater for subsequent RNA analysis. Short‐term storage and shipment of well‐preserved tumor tissue are clearly feasible for all institutions, thereby facilitating large multiinstitutional studies of biological prognostic factors. Med. Pediatr. Oncol. 34:438–442, 2000. © 2000 Wiley‐Liss, Inc.

Gene Expression in Temporal Lobe Epilepsy is Consistent with Increased Release of Glutamate by Astrocytes

“…RNAlater has been shown to preserve RNA in tissue samples at least as well, if not better than, rapid freezing in liquid nitrogen (20). The hippocampus was microdissected into subregions and total RNA extracted from the CA1 subregion using the RNeasy kit for lipid tissue (Qiagen, Valencia, CA.).…”

Aspects of Quality and Project Management in Analyses of Large Scale Sequencing Data

“…It is a non toxic, non flammable liquid that can be transported everywhere without any problems (even in airplanes) and it preserves RNA at room temperature at least for 5-7 days (Grotzer et al, 2000; product descriptions of e.g. Qiagen, Applied Biosystems) without loss of quality compared to frozen samples (Grotzer et al, 2000;Mutter et al, 2004;Gorokova, 2005). A closed cooling chain is not mandatory.…”

Utilização do produto Allprotect Tissue Reagent® na estabilização do DNA extraído de tecidos dentais humanos em diferentes condições de armazenamento

“…No entanto, sabe-se que algumas situações podem dificultar a manutenção das amostras em condições adequadas e em baixas temperaturas, como nos casos em que o acesso ao local com os materiais necessários não é possível (Grotzer et al, 2000).…”

“…Nesse contexto, a utilização de produtos que estabilizam as moléculas de DNA seria uma alternativa para a manutenção da qualidade da amostra até que a mesma possa ser transportada para os laboratórios competentes (Grotzer et al, 2000). Nesse sentido, testes envolvendo a eficiência desses produtos devem ser realizados na tentativa de justificar o seu uso.…”

“…Assim, torna-se possível que as evidências recolhidas em qualquer lugar à temperatura ambiente sejam enviadas para as instalações apropriadas de forma segura (Grotzer et al, 2000;Medeiros et al, 2003;Mutter et al, 2004;Qiagen, 2006Qiagen, , 2008Qiagen, , 2011 (Mee et al, 2011;Gelbart et al, 2012), ainda não há trabalhos na literatura que evidenciam a funcionalidade e eficácia do mesmo em relação aos tecidos dentais.…”