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使用低成本的NAxtra磁性纳米颗粒从少量或单个细胞中快速分离DNA和RNA。

Rapid DNA and RNA isolation from few or single cells using low-cost NAxtra magnetic nanoparticles.

作者信息

Starheim Eirin Johannessen, Manaf Adeel, Hashim Adnan, Andersen Niklas Nonboe, Ravlo Erlend, Wang Wei, Saasen Vidar Langseth, Liabakk Nina-Beate, Erlandsen Sten Even, Aas Per Arne, Hagen Lars, Bjørås Magnar

机构信息

Department of Clinical and Molecular Medicine (IKOM), Norwegian University of Science and Technology (NTNU), 7491, Trondheim, Norway.

Department of Microbiology, Oslo University Hospital and University of Oslo, 0372, Oslo, Norway.

出版信息

Sci Rep. 2025 Jul 2;15(1):22472. doi: 10.1038/s41598-025-05770-y.

Abstract

A novel, cost-effective nucleic acid (NA) isolation method for purifying total NA, DNA, or RNA from both two- and three-dimensional cell cultures has been developed at the Norwegian University of Science and Technology utilizing NAxtra magnetic nanoparticles. This method achieves comparable yields to existing isolation kits while offering significant improvements in cost and processing speed. However, the original protocol was not optimized for small-cell numbers or single-cell applications. Given the growing interest in single-cell and rare-cell population studies, there is a critical need for more sensitive isolation techniques. In this study, we have enhanced the sensitivity of the NAxtra-based isolation method to facilitate mid- to high-throughput purification from as few as 10,000 cells down to single cells. Automated processing using KingFisher systems enables the rapid handling of 96 samples within 12-18 min. Our findings indicate that this method not only matches but can exceed the performance of existing alternatives in (RT)-qPCR detection while being significantly more economical and efficient. Additionally, it enables the extraction of high-quality RNA suitable for transcriptomics analyses from limited cell quantities, including single cells. This advancement holds substantial promise for improving the accessibility and efficiency of NA research, particularly in studies involving scarce cellular materials.

摘要

挪威科技大学利用NAxtra磁性纳米颗粒开发了一种新颖且经济高效的核酸(NA)分离方法,用于从二维和三维细胞培养物中纯化总NA、DNA或RNA。该方法在产量上与现有分离试剂盒相当,同时在成本和处理速度方面有显著提升。然而,原始方案并未针对少量细胞或单细胞应用进行优化。鉴于对单细胞和稀有细胞群体研究的兴趣日益浓厚,迫切需要更灵敏的分离技术。在本研究中,我们提高了基于NAxtra的分离方法的灵敏度,以促进从中高通量地从低至10000个细胞直至单个细胞进行纯化。使用KingFisher系统进行自动化处理能够在12 - 18分钟内快速处理96个样品。我们的研究结果表明,该方法不仅在(RT)-qPCR检测中与现有替代方法性能相当,甚至可以超越,而且更加经济高效。此外,它能够从有限数量的细胞(包括单个细胞)中提取适用于转录组学分析的高质量RNA。这一进展对于提高NA研究的可及性和效率具有重要意义,特别是在涉及稀缺细胞材料的研究中。

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