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一种高效且可回收的电洗脱装置:从自制到模块化设计,有望实现大规模生产。

An efficient and recyclable electroeluter: from homemade to modular design for potential mass production.

机构信息

College of Chemistry, Jilin University, 130012 Changchun, People's Republic of China.

出版信息

Lab Chip. 2023 Aug 22;23(17):3874-3881. doi: 10.1039/d3lc00428g.

Abstract

Electrophoresis is one of the most powerful techniques to separate nucleic acids or protein molecules. The recovery of purified components from the gel is key to downstream analysis or function study. Here, we provide a cost-effective electroeluter in both homemade and module-assembled versions. The recovery yield can reach as high as >90% for single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), and protein in practical testing, which outperforms a commercial kit as well as a purchased electroeluter. It fully addresses the existing concerns in this field. First of all, for almost all kits, there remains ambiguity in recovering ssDNA to satisfy specific demands, which is generally ignored. Secondly, the recovery of dsDNA from agarose gel with consumables is vulnerable to a lot of factors and involves chemicals/materials that are not friendly to the environment and operating personnel. Thirdly, recovery from polyacrylamide matrices is very difficult, and the most exploited diffusion method through crush-and-soak suffers from low yield even after a long-time diffusion. Lastly, there is a universal problem in scaling up, especially for commercial electroeluters. The present electroelution method addresses the above issues, and it is believed that it will facilitate associated research and find widespread application.

摘要

电泳是分离核酸或蛋白质分子的最强大技术之一。从凝胶中回收纯化的成分是下游分析或功能研究的关键。在这里,我们提供了一种具有成本效益的电洗脱器,有自制和模块组装两种版本。在实际测试中,该电洗脱器对单链 DNA(ssDNA)、双链 DNA(dsDNA)和蛋白质的回收率高达>90%,优于商业试剂盒和购买的电洗脱器。它完全解决了该领域现有的问题。首先,对于几乎所有的试剂盒来说,在满足特定需求方面,回收 ssDNA 仍然存在模糊性,这通常被忽略了。其次,使用耗材从琼脂糖凝胶中回收 dsDNA 容易受到许多因素的影响,并且涉及到对环境和操作人员不友好的化学品/材料。第三,从聚丙烯酰胺基质中回收非常困难,最常用的通过粉碎和浸泡的扩散方法即使经过长时间的扩散也会导致产量低。最后,规模化存在普遍的问题,特别是对于商业电洗脱器。本电洗脱方法解决了上述问题,相信它将促进相关研究,并得到广泛应用。

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