Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation
Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation
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DOI：
10.1371/journal.pbio.3000107
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发表时间：
2019-01-01
期刊：
影响因子：
9.8
通讯作者：
Jurkowski, Tomasz P.
中科院分区：
文献类型：
--
作者：
Oberacker, Phil;Stepper, Peter;Jurkowski, Tomasz P.
目前的分子生物学实验室严重依赖于核酸的纯化和操作。然而，常用的基于离心机和柱的方案需要专门的设备，通常使用有毒试剂，并且在经济上不可扩展或不实用于以高通量方式使用。尽管人们已经知道磁珠可以为这些问题提供一个优雅的答案，但基于磁珠的开源协议的开发一直受到限制。在这篇文章中，我们提供了一个简单的功能化磁珠的合成一步一步的说明，并详细的协议，他们在高通量纯化质粒，基因组DNA，RNA和总核酸（TNA）从一系列的细菌，动物，植物，环境和合成来源的使用。我们还提供了一个基于珠的方案，用于亚硫酸氢盐转化和DNA和RNA片段的大小选择。与其他方法的比较突出了使用磁珠的能力，多功能性和极高的成本效益。这些开源协议和相关网页（bomb.bio）可以作为进一步协议定制和社区参与的平台。
Current molecular biology laboratories rely heavily on the purification and manipulation of nucleic acids. Yet, commonly used centrifuge-and column-based protocols require specialised equipment, often use toxic reagents, and are not economically scalable or practical to use in a high-throughput manner. Although it has been known for some time that magnetic beads can provide an elegant answer to these issues, the development of open-source protocols based on beads has been limited. In this article, we provide step-by-step instructions for an easy synthesis of functionalised magnetic beads, and detailed protocols for their use in the high-throughput purification of plasmids, genomic DNA, RNA and total nucleic acid (TNA) from a range of bacterial, animal, plant, environmental and synthetic sources. We also provide a bead-based protocol for bisulfite conversion and size selection of DNA and RNA fragments. Comparison to other methods highlights the capability, versatility, and extreme cost-effectiveness of using magnetic beads. These open-source protocols and the associated webpage (https://bomb.bio) can serve as a platform for further protocol customisation and community engagement.