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An efficient method for long-term room temperature storage of RNA

机译:RNA长期室温保存的有效方法

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摘要

RNA is a tool used in many fields, from molecular and cellular biology to medicine and nanotechnology. For most of these uses, the integrity of RNA is required and must be maintained during storage. Even though freezing is currently the storage method of choice, the increasing number of samples to be stored and the costly use of a cold chain have highlighted the need for room temperature preservation methods. Here, we report a new room temperature technology that consists in drying RNA samples in the presence of a stabilizer in stainless steel minicapsules. These air- and water-tight capsules isolate RNA from the atmosphere and maintain an anhydrous and anoxic environment. Through the evaluation of RNA integrity over time at room temperature or 90 °C, we identified atmospheric humidity as a major deleterious factor. The degradation rate dependence in temperature fitted an Arrhenius model, with an activation energy of 28.5 kcal/mol and an extrapolated room temperature degradation rate of 3.2 10−13t/s (95% confidence interval: 2.3–4.2t/s). In these conditions, it is expected that an RNA molecule will be subjected to 0.7–1.3 cut every 1000 nucleotides per century. In addition, we showed that stored RNA is compatible for further analyses, such as reverse transcription-quantitative PCR. No significant change in the Cq values was observed over a simulated period of several decades. At last, our data are consistent with a sequence-independent degradation rate of RNA in the solid state.
机译:RNA是从分子和细胞生物学到医学和纳米技术的许多领域中使用的工具。对于大多数这些用途,RNA的完整性是必需的,必须在存储过程中保持。尽管目前选择冷冻是一种存储方法,但要存储的样品数量不断增加以及冷链的昂贵使用,凸显了对室温保存方法的需求。在这里,我们报告了一种新的室温技术,该技术包括在不锈钢微胶囊中存在稳定剂的情况下干燥RNA样品。这些气密和水密的胶囊将RNA与大气隔离,并保持无水缺氧的环境。通过评估室温或90°C下随时间变化的RNA完整性,我们确定了大气湿度是主要的有害因素。温度对温度的降解率依赖性符合Arrhenius模型,其活化能为28.5 kcal / mol,外推的室温降解率为3.2 10 −13 / nt / s(95%置信区间:2.3) –4.2 / nt / s)。在这些条件下,预计一个世纪每一个1000个核苷酸将对RNA分子进行0.7-1.3切割。此外,我们证明了所存储的RNA可用于进一步的分析,例如逆转录定量PCR。在几十年的模拟时间内,未观察到Cq值的显着变化。最后,我们的数据与固态RNA的序列独立降解速率一致。

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