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Application of short monolithic columns for fast purification of plasmid DNA

机译:短整体柱在质粒DNA快速纯化中的应用

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Anion-exchange chromatography is one of the most important methods in downstream processing of plasmid DNA, both as a process and as an analytical technique. Separation of plasmid DNA on traditional particle-based anion-exchange supports is usually slow. Moreover, such supports have a low capacity for plasmid DNA due to the steric exclusion effects. In this work, the separation of plasmid DNA using short monolithic columns, Convective Interaction Media, will be presented. It will be demonstrated that plasmid DNA can be purified from bacterial cells using alkaline lysis followed by chromatography on a very short weak anion-exchange chromatographic columns-disks-with good purity and quality within a short time. Furthermore, the separation of plasmid DNA from cell RNA can be carried out without the need of adding RNAse. Fast and efficient method for in-process control of the purified plasmid will be described as well.
机译:阴离子交换色谱是质粒DNA下游处理中最重要的方法之一,无论是作为一种方法还是作为一种分析技术。在传统的基于颗粒的阴离子交换载体上分离质粒DNA通常很慢。而且,由于空间排阻作用,这种支持物对质粒DNA的能力低。在这项工作中,将介绍使用短的整体柱(对流相互作用介质)分离质粒DNA的方法。可以证明,可以使用碱裂解法从细菌细胞中纯化质粒DNA,然后在很短的时间内在极短的弱阴离子交换色谱柱-磁盘上进行色谱分离,从而获得良好的纯度和质量。此外,可以从细胞RNA中分离质粒DNA,而无需添加RNAse。也将描述用于纯化质粒的过程中控制的快速和有效方法。

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