Clermont Dominique, Santoni Sylvain, Saker Safa, Gomard Maite, Gardais Eliane, Bizet Chantal
1 Institut Pasteur, Collection de l'Institut Pasteur , Département de Microbiologie, Paris, France .
Biopreserv Biobank. 2014 Jun;12(3):176-83. doi: 10.1089/bio.2013.0082.
A new procedure for room-temperature storage of DNA was evaluated whereby DNA samples from human tissue, bacteria, and plants were stored under an anoxic and anhydrous atmosphere in small glass vials fitted in stainless-steel, laser-sealed capsules (DNAshells(®)). Samples were stored in DNAshells(®) at room temperature for various periods of time to assess any degradation and compare it to frozen control samples and those stored in GenTegra™ tubes. The study included analysis of the effect of accelerated aging by using a high temperature (76°C) at 50% relative humidity. No detectable DNA degradation was seen in samples stored in DNAshells(®) at room temperature for 18 months. Polymerase chain reaction experiments, pulsed field gel electrophoresis, and amplified fragment length polymorphism analyses also demonstrated that the protective properties of DNAshells(®) are not affected by storage under extreme conditions (76°C, 50% humidity) for 30 hours, guaranteeing 100 years without DNA sample degradation. However, after 30 hours of storage at 76°C, it was necessary to include adjustments to the process in order to avoid DNA loss. Successful protection of DNA was obtained for 1 week and even 1 month of storage at high temperature by adding trehalose, which provides a protective matrix. This study demonstrates the many advantages of using DNAshells(®) for room-temperature storage, particularly in terms of long-term stability, safety, transport, and applications for molecular biology research.
对一种DNA室温储存的新方法进行了评估,该方法是将来自人体组织、细菌和植物的DNA样本在无氧无水的气氛下储存在安装在不锈钢激光密封胶囊(DNAshells(®))中的小玻璃瓶中。将样本在DNAshells(®)中于室温下储存不同时间,以评估是否有降解,并将其与冷冻对照样本以及储存在GenTegra™管中的样本进行比较。该研究包括通过在50%相对湿度下使用高温(76°C)来分析加速老化的影响。在室温下于DNAshells(®)中储存18个月的样本未检测到DNA降解。聚合酶链反应实验、脉冲场凝胶电泳和扩增片段长度多态性分析也表明,DNAshells(®)的保护性能不受在极端条件(76°C,50%湿度)下储存30小时的影响,保证100年DNA样本不降解。然而,在76°C储存30小时后,有必要对该过程进行调整以避免DNA损失。通过添加提供保护基质的海藻糖,在高温下成功保护DNA达1周甚至1个月。这项研究证明了使用DNAshells(®)进行室温储存的诸多优点,特别是在长期稳定性、安全性、运输以及分子生物学研究应用方面。