Oberacker Phil, Stepper Peter, Bond Donna, Hipp Katharina, Hore Timothy A, Jurkowski Tomasz P
Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
School of Biosciences, Cardiff University, Cardiff, Wales, United Kingdom.
Bio Protoc. 2019 Oct 20;9(20):e3394. doi: 10.21769/BioProtoc.3394.
The purification of nucleic acids is one of the most common procedures employed in modern molecular biology laboratories. Typically, commercial column-based protocols are utilized to isolate DNA or RNA from various sources. However, these methods not only require specialized equipment, but are also extremely expensive for high-throughput applications. Although an elegant answer to this issue can be provided by paramagnetic beads, bead-based open-source protocols have been limited in the past. Here, we provide an easy to follow step-by-step manual for the synthesis of paramagnetic beads, as well as their functionalization with either a silica- or a carboxyl-surface that can be used to replace the commercial columns with self-made magnetic beads. Together with a variety of detailed protocols for their use in high-throughput nucleic acids extractions, this bead synthesis method forms the recently published open platform Bio-On-Magnetic-Beads (BOMB), which is available on PLOS Biology ( Oberacker , 2019 ). Updated protocols can be found on the associated webpage (https://bomb.bio).
核酸纯化是现代分子生物学实验室中最常用的操作程序之一。通常，基于商业柱的方案被用于从各种来源分离DNA或RNA。然而，这些方法不仅需要专门的设备，而且对于高通量应用来说成本极高。尽管顺磁珠可以很好地解决这个问题，但过去基于珠子的开源方案一直很有限。在这里，我们提供了一个易于遵循的顺磁珠合成分步手册，以及用二氧化硅或羧基表面对其进行功能化的方法，这些顺磁珠可用于用自制磁珠替代商业柱。连同各种关于其在高通量核酸提取中的详细使用方案，这种珠子合成方法构成了最近发表的开放平台生物磁珠（Bio-On-Magnetic-Beads，BOMB），该平台可在《公共科学图书馆·生物学》（Oberacker，2019年）上获取。更新后的方案可在相关网页（https://bomb.bio）上找到。
