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Scalable Nucleic Acid Storage and Retrieval Using Barcoded Microcapsules
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2021-10-15 , DOI: 10.1021/acsami.1c14985
James L Banal 1 , Mark Bathe 1, 2
Affiliation  

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.

中文翻译:

使用条形码微胶囊进行可扩展的核酸存储和检索

核酸测序和合成技术的快速发展激发了从病毒、细菌和哺乳动物来源和生物体中收集、储存和测序 DNA 和 RNA 的主要需求。然而,目前存储核酸的方法依赖于低温环境,并且需要机器人来访问,这对可扩展和低成本的核酸存储提出了挑战。在这里,我们提出了一种存储核酸的替代方法,称为使用 barcOded micRocCapsules (PANDORA) 保存和获取核酸。跨越千碱基到千兆碱基以及来自不同来源(包括动物、细菌和病毒)的核酸被封装在二氧化硅微胶囊中,以保护它们在室温下免受环境变性剂的影响。附着在每个微胶囊上的分子条形码可以使用荧光激活分选进行样本汇集和随后的识别和检索。我们展示了从池中定量存储和快速访问目标核酸,模拟在传统存储系统中实施的标准检索操作,包括恢复 100,000-200,000 个样本和使用四个唯一条形码的布尔逻辑选择。对检索到的样本进行定量聚合酶链反应和短读长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具有高样品保真度的可扩展的长期、室温存储和检索核酸。我们展示了从池中定量存储和快速访问目标核酸,模拟在传统存储系统中实施的标准检索操作,包括恢复 100,000-200,000 个样本和使用四个唯一条形码的布尔逻辑选择。对检索到的样本进行定量聚合酶链反应和短读长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具有高样品保真度的可扩展的长期、室温存储和检索核酸。我们展示了从池中定量存储和快速访问目标核酸,模拟在传统存储系统中实施的标准检索操作,包括恢复 100,000-200,000 个样本和使用四个唯一条形码的布尔逻辑选择。对检索到的样本进行定量聚合酶链反应和短读长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具有高样品保真度的可扩展的长期、室温存储和检索核酸。对检索到的样本进行定量聚合酶链反应和短读长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具有高样品保真度的可扩展的长期、室温存储和检索核酸。对检索到的样本进行定量聚合酶链反应和短读长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具有高样品保真度的可扩展的长期、室温存储和检索核酸。
更新日期:2021-10-27
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