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Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits

机译:通过PCR纯化和凝胶提取试剂盒快速再生和重用硅胶柱

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摘要

Silica columns from PCR purification and gel extraction kits are widely used in laboratories worldwide to assist in gene cloning. However, the use of these columns can generate plastic waste that has an environmental impact due to their one-off design and massive consumption. Thus, it is important to develop a novel method that can reduce the utilization of silica columns but not affect research efficiency. In this study, various chemical and nonchemical reagents were used to eliminate residual DNA within used columns from PCR purification and gel extraction kits. We show that phosphoric acid is the most effective reagent among those tested to remove DNA contamination from used columns. Columns regenerated using 1 M phosphoric acid have a DNA purification capability that is comparable to that of fresh columns. We demonstrate that silica columns can be regenerated and reused a minimum of five times. The lab-made buffers are compatible with the regenerated columns for DNA purification, and DNA that is prepared with the regenerated columns can be used for gene cloning without affecting the gene cloning efficiency. Thus, the use of this novel method greatly reduces the production of laboratory waste and benefits numerous laboratories worldwide.
机译:PCR纯化和凝胶提取试剂盒中的硅胶柱在全世界的实验室中广泛使用,以协助基因克隆。但是,由于这些色谱柱的一次性设计和大量消耗,使用这些色谱柱会产生塑料废物,对环境产生影响。因此,重要的是开发一种可以减少硅胶柱利用率但又不影响研究效率的新方法。在这项研究中,使用了各种化学和非化学试剂来消除PCR纯化和凝胶提取试剂盒中所用色谱柱中的残留DNA。我们表明,磷酸是在从使用过的色谱柱中去除DNA污染的那些试剂中最有效的试剂。使用1 M磷酸再生的色谱柱具有与新鲜色谱柱相当的DNA纯化能力。我们证明了硅胶柱可以再生和至少重复使用五次。实验室制备的缓冲液与用于DNA纯化的再生柱兼容,并且使用再生柱制备的DNA可以用于基因克隆而不会影响基因克隆效率。因此,使用这种新颖的方法极大地减少了实验室废物的产生,并使全世界许多实验室受益。

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