Laboratoire de Recherche et développement, Imagene Company, Pessac, France.
Imagene, plateforme de production, Genopole, Evry, France.
PLoS One. 2021 Nov 11;16(11):e0259868. doi: 10.1371/journal.pone.0259868. eCollection 2021.
DNA conservation is central to many applications. This leads to an ever-increasing number of samples which are more and more difficult and costly to store or transport. A way to alleviate this problem is to develop procedures for storing samples at room temperature while maintaining their stability. A variety of commercial systems have been proposed but they fail to completely protect DNA from deleterious factors, mainly water. On the other side, Imagene company has developed a procedure for long-term conservation of biospecimen at room temperature based on the confinement of the samples under an anhydrous and anoxic atmosphere maintained inside hermetic capsules. The procedure has been validated by us and others for purified RNA, and for DNA in buffy coat or white blood cells lysates, but a precise determination of purified DNA stability is still lacking. We used the Arrhenius law to determine the DNA degradation rate at room temperature. We found that extrapolation to 25°C gave a degradation rate constant equivalent to about 1 cut/century/100 000 nucleotides, a stability several orders of magnitude larger than the current commercialized processes. Such a stability is fundamental for many applications such as the preservation of very large DNA molecules (particularly interesting in the context of genome sequencing) or oligonucleotides for DNA data storage. Capsules are also well suited for this latter application because of their high capacity. One can calculate that the 64 zettabytes of data produced in 2020 could be stored, standalone, for centuries, in about 20 kg of capsules.
DNA 保守性在许多应用中至关重要。这导致越来越多的样本难以存储或运输,而且成本越来越高。缓解这个问题的一种方法是开发在室温下存储样本的程序,同时保持其稳定性。已经提出了各种商业系统,但它们不能完全防止 DNA 受到有害因素(主要是水)的影响。另一方面,Imagene 公司开发了一种在室温下长期保存生物样本的程序,该程序基于将样本限制在密封胶囊内的无水和缺氧气氛中。该程序已经被我们和其他人验证用于纯化的 RNA,以及用于 buffy 涂层或白细胞裂解物中的 DNA,但对纯化 DNA 稳定性的精确测定仍然缺乏。我们使用阿仑尼乌斯定律来确定 DNA 在室温下的降解速率。我们发现,外推至 25°C 得到的降解速率常数相当于每 100000 个核苷酸约 1 个切割/世纪,稳定性比当前商业化的过程大几个数量级。这种稳定性对于许多应用非常重要,例如非常大的 DNA 分子的保存(特别是在基因组测序的背景下)或用于 DNA 数据存储的寡核苷酸。由于其高容量,胶囊也非常适合后者的应用。可以计算,2020 年产生的 64ZB 数据,如果单独存储,可以在大约 20 公斤的胶囊中保存数百年。