The Filterprep: A simple and efficient approach for high-yield, high-purity plasmid DNA purification

IF 4.9 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Yu-Hsuan Cheng , Yu-Jiu Wu , Yung-Chun Shih , Yu-Qian Lin , Yu-Wei Chang , Yu-Heng Wu , En-Wei Hu , Ren-Hsuan Ku , Cheng-Mu Wu , Shao-Chi Wu , Ting-Yu Yeh , Chung-Te Chang
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引用次数: 0

Abstract

Plasmid DNA purification remains a fundamental yet resource-intensive step in molecular biology and biotechnology. Conventional protocols often yield plasmids with suboptimal purity, while commercial kits, though efficient, are costly and use proprietary formulations with limited transparency for troubleshooting or customization. Here, we present Filterprep, a hybrid method combining classical ethanol precipitation with a single spin-column cleanup step, enabling rapid recovery of high-purity plasmid DNA with yields notably higher than those obtained using commercial kits, in approximately 40 min, with yields up to 5-fold greater than those of representative commercial midiprep kits under matched conditions. Filterprep offers a transparent, cost-effective, and scalable alternative, simplifying workflows and reducing hands-on time without compromising DNA quality. The purified plasmids are compatible with a variety of downstream applications, including mammalian cell transfection, protein expression, and reporter assays, while remaining stable for 12 months at −20 °C. With its balance of efficiency and practicality, Filterprep is well suited for high-throughput laboratories and resource-limited settings.
Filterprep:一种简单有效的高产、高纯度质粒DNA纯化方法
质粒DNA纯化仍然是分子生物学和生物技术中一个基础但资源密集的步骤。传统方法通常产生纯度不理想的质粒,而商业试剂盒虽然有效,但价格昂贵,并且使用专有配方,对故障排除或定制的透明度有限。在这里,我们提出了Filterprep,这是一种结合了经典乙醇沉淀和单个自旋柱清理步骤的混合方法,能够快速回收高纯度质粒DNA,产量明显高于使用商业试剂盒获得的产量,大约40 min,产量比具有代表性的商业中等准备试剂盒在匹配条件下的产量高出5倍。Filterprep提供了一种透明,经济高效,可扩展的替代方案,简化了工作流程,减少了动手时间,而不影响DNA质量。纯化的质粒与多种下游应用兼容,包括哺乳动物细胞转染,蛋白质表达和报告基因检测,同时在- 20°C下保持稳定12个月。凭借其效率和实用性的平衡,Filterprep非常适合高通量实验室和资源有限的设置。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
New biotechnology
New biotechnology 生物-生化研究方法
CiteScore
11.40
自引率
1.90%
发文量
77
审稿时长
1 months
期刊介绍: New Biotechnology is the official journal of the European Federation of Biotechnology (EFB) and is published bimonthly. It covers both the science of biotechnology and its surrounding political, business and financial milieu. The journal publishes peer-reviewed basic research papers, authoritative reviews, feature articles and opinions in all areas of biotechnology. It reflects the full diversity of current biotechnology science, particularly those advances in research and practice that open opportunities for exploitation of knowledge, commercially or otherwise, together with news, discussion and comment on broader issues of general interest and concern. The outlook is fully international. The scope of the journal includes the research, industrial and commercial aspects of biotechnology, in areas such as: Healthcare and Pharmaceuticals; Food and Agriculture; Biofuels; Genetic Engineering and Molecular Biology; Genomics and Synthetic Biology; Nanotechnology; Environment and Biodiversity; Biocatalysis; Bioremediation; Process engineering.
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