Dipartimento di Scienza e Tecnologia del Farmaco, Università di Torino, Via P. Giuria 9, 10125, Turin, Italy.
Department of Chemistry, Iowa State University, 1605 Gilman Hall, Ames, IA, 50011, USA.
Anal Bioanal Chem. 2019 Oct;411(25):6583-6590. doi: 10.1007/s00216-019-02054-z. Epub 2019 Aug 17.
This study reports a follow-up investigation on the capture of specific DNA sequences using ion-tagged oligonucleotides (ITOs) and magnetic ionic liquids (MIL). Five allylimidazolium salts bearing octyl substituents ([AOIM]-ITOs) were used for the selective extraction of the internal transcribed spacer region (ITS) from Arabidopsis thaliana. In this work, the ability of the [AOIM]-ITOs to enhance the extraction of longer target sequences (~ 700 bp) of plant origin was shown. Moreover, the independence of the probe binding position and the importance of complementarity to the target region for the extraction performance were demonstrated. To test the specificity of the ITOs, the same experiments were performed using the ITS region from another plant species, with a lower target capture for the probes which were specific for the A. thaliana sequence. Finally, extraction in the presence of interferences (heterogenous DNA, primary and secondary metabolites, proteins) provided interesting and insightful results. This work illustrates the feasibility and versatility of these probes when coupled to MILs for rapid, cost-effective, and environmentally sensitive sample preparation in the extraction of specific target sequences from different origins. Graphical abstract.
本研究报告了使用离子标记寡核苷酸（ITOs）和磁性离子液体（MIL）捕获特定 DNA 序列的后续调查。五种带有辛基取代基的烯丙基咪唑鎓盐（[AOIM]-ITOs）被用于从拟南芥中选择性提取内部转录间隔区（ITS）。在这项工作中，[AOIM]-ITOs 增强了对更长目标序列（~700 bp）的提取能力。此外，还证明了探针结合位置的独立性和与目标区域互补性对提取性能的重要性。为了测试 ITOs 的特异性，使用另一种植物物种的 ITS 区域进行了相同的实验，对于特异性针对 A. thaliana 序列的探针，目标捕获率较低。最后，在存在干扰物（异源 DNA、初级和次级代谢物、蛋白质）的情况下进行提取，提供了有趣和有见地的结果。这项工作说明了这些探针与 MIL 结合用于从不同来源快速、经济高效且环境敏感地提取特定目标序列的样品制备的可行性和多功能性。
