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基于微珠的DNA合成与测序,用于利用数字微流控技术进行集成数据存储

Bead-Based DNA Synthesis and Sequencing for Integrated Data Storage Using Digital Microfluidics.

作者信息

Piao Yuhao, Fang Yitong, Li Bin, Man Tiantian, Chen Jie, Zhu Fulin, Wang Weiqiang, Wan Ying, Deng Shengyuan

机构信息

School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, 210094, China.

School of Mechanical Engineering, Nanjing University of Science and Technology, Nanjing, 210094, China.

出版信息

Angew Chem Int Ed Engl. 2025 Jan 21;64(4):e202416004. doi: 10.1002/anie.202416004. Epub 2024 Dec 5.

DOI:10.1002/anie.202416004
PMID:39606901
Abstract

DNA is considered as a prospective candidate for the next-generation data storage medium, due to its high coding density, long cold-storage lifespan, and low energy consumption. Despite these advantages, challenges remain in achieving high-fidelity, fully integrated, and cost-efficient DNA storage system. In this study, a homemade digital microfluidic (DMF)-based compact DNA data storing pipeline is orchestrated to complete the entire process from the synthesis to the sequencing. The synthetic half employs phosphoramidite chemistry on 200 nm magnetic beads (MBs), where the dimethyltrityl protecting group is removed by droplet manipulation of trichloroacetic acid. The sequencing counterpart relies on pyrophosphate releasing originated from polymerase-catalyzed primer extension, which leads to photon-countable chemiluminescence (CL) signal in 2.5-μL drops of trienzyme cascading reactions. Further by DNA denaturation, repeated pyrosequencing plus plurality voting can improve the nucleobase accuracy beyond 95 %. As a proof-of-concept trial, semantic information is saved in DNA via the Huffman coding algorithm plus the Reed-Solomon error-correction, and then robustly retrieved from this streamlined platform. As a result, it took a net total of approximately 6.5 h to writing and reading 8 bytes of data, that equal to a storaging speed of 49 min/byte, much quicker than the previously reported 2.8-4.2 h/byte. This bead-based miniaturized device promises an unattended protocol for achieving high-throughput, full-packaged, and above all, neatly precision DNA storage.

摘要

由于DNA具有高编码密度、长冷存储寿命和低能耗等特点,它被认为是下一代数据存储介质的潜在候选者。尽管有这些优点,但在实现高保真、完全集成且具有成本效益的DNA存储系统方面仍存在挑战。在本研究中,精心设计了一种基于自制数字微流控(DMF)的紧凑型DNA数据存储流程,以完成从合成到测序的整个过程。合成部分在200 nm磁珠(MBs)上采用亚磷酰胺化学方法,通过三氯乙酸的液滴操作去除二甲基三苯甲基保护基团。测序部分则依赖于聚合酶催化引物延伸产生的焦磷酸释放,这在2.5 μL三酶级联反应液滴中产生可光子计数的化学发光(CL)信号。进一步通过DNA变性、重复焦磷酸测序和多数投票,可以将碱基准确率提高到95%以上。作为概念验证试验,通过霍夫曼编码算法和里德 - 所罗门纠错将语义信息存储在DNA中,然后从这个简化平台中可靠地检索出来。结果,写入和读取8字节数据总共花费约6.5小时,相当于存储速度为49分钟/字节,比之前报道的2.8 - 4.2小时/字节快得多。这种基于磁珠的小型化设备有望实现一种无人值守的协议,以实现高通量、全封装且最重要的是精确的DNA存储。

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