Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 United States.
Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142 United States.
ACS Appl Mater Interfaces. 2021 Oct 27;13(42):49729-49736. doi: 10.1021/acsami.1c14985. Epub 2021 Oct 15.
Rapid advances in nucleic acid sequencing and synthesis technologies have spurred a major need to collect, store, and sequence the DNA and RNA from viral, bacterial, and mammalian sources and organisms. However, current approaches to storing nucleic acids rely on a low-temperature environment and require robotics for access, posing challenges for scalable and low-cost nucleic acid storage. Here, we present an alternative method for storing nucleic acids, termed Preservation and Access of Nucleic aciDs using barcOded micRocApsules (PANDORA). Nucleic acids spanning kilobases to gigabases and from different sources, including animals, bacteria, and viruses, are encapsulated into silica microcapsules to protect them from environmental denaturants at room temperature. Molecular barcodes attached to each microcapsule enable sample pooling and subsequent identification and retrieval using fluorescence-activated sorting. We demonstrate quantitative storage and rapid access to targeted nucleic acids from a pool emulating standard retrieval operations implemented in conventional storage systems, including recovery of 100,000-200,000 samples and Boolean logic selection using four unique barcodes. Quantitative polymerase chain reaction and short-read sequencing of the retrieved samples validated the sorting experiments and the integrity of the released nucleic acids. Our proposed approach offers a scalable long-term, room-temperature storage and retrieval of nucleic acids with high sample fidelity.
核酸测序和合成技术的快速发展,极大地推动了从病毒、细菌和哺乳动物来源和生物中收集、存储和测序 DNA 和 RNA 的需求。然而,目前储存核酸的方法依赖于低温环境,并且需要机器人进行访问,这给可扩展和低成本的核酸存储带来了挑战。在这里,我们提出了一种储存核酸的替代方法,称为使用条形码微胶囊保存和获取核酸(PANDORA)。跨越千碱基到千兆碱基的核酸,以及来自不同来源的核酸,包括动物、细菌和病毒,都被封装到二氧化硅微胶囊中,以在室温下保护它们免受环境变性剂的影响。每个微胶囊上附加的分子条形码可实现样品的混合,并随后使用荧光激活分选进行鉴定和检索。我们展示了从模拟传统存储系统中实施的标准检索操作的样本池中定量存储和快速获取靶向核酸的能力,包括回收 100,000-200,000 个样本,以及使用四个独特条形码进行布尔逻辑选择。回收样本的定量聚合酶链反应和短读测序验证了分选实验和释放核酸的完整性。我们提出的方法提供了一种可扩展的长期、室温下储存和检索核酸的方法,具有高样品保真度。