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Extraction of DNA by Magnetic Ionic Liquids: Tunable Solvents for Rapid and Selective DNA Analysis

机译:磁性离子液体提取DNA:用于快速和选择性DNA分析的可调溶剂

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DNA extraction represents a significant bottleneck in nucleic acid analysis. In this study, hydrophobic magnetic ionic liquids (MILs) were synthesized and employed as solvents for the rapid and efficient extraction of DNA from aqueous solution. The DNA-enriched microdroplets were manipulated by application of a magnetic field. The three MILs examined in this study exhibited unique DNA extraction capabilities when applied toward a variety of DNA samples and matrices. High extraction efficiencies were obtained for smaller single-stranded and double-stranded DNA using the benzyltrioctylammonium bromotrichloroferrate(III) ([(C-8)(3)BnN+][FeCl3Br-]) MIL, while the dicationic 1,12-di(3-hexadecylbenzimidazolium)dodecane bis[(trifluoromethyl)sulfonyl]imide bromotrichloroferrate(III) ([(C16BnIM)(2)C-122(+)][NTf2-, FeCl3Br-]) MIL produced higher extraction efficiencies for larger DNA molecules. The MIL-based method was also employed for the extraction of DNA from a complex matrix containing albumin, revealing a competitive extraction behavior for the trihexyl(tetradecyl)phosphonium tetrachloroferrate(III) ([P-6,6,6,14(+)][FeCl4-]) MIL in contrast to the [(C-8)(BnN+)-Bn-3][FeCl3Br-] MIL, which resulted in significantly less coextraction of albumin. The MIL-DNA method was employed for the extraction of plasmid DNA from bacterial cell lysate. DNA of sufficient quality and quantity for polymerase chain reaction (PCR) amplification was recovered from the MIL extraction phase, demonstrating the feasibility of MIL-based DNA sample preparation prior to downstream analysis.
机译:DNA提取代表了核酸分析中的重大瓶颈。在这项研究中,合成了疏水性磁性离子液体(MILs),并将其用作从水溶液中快速有效提取DNA的溶剂。通过施加磁场来操纵富含DNA的微滴。在本研究中检测的三个MIL应用于多种DNA样品和基质时,具有独特的DNA提取能力。使用苄基三辛基溴化三氯高铁酸盐(III)([(C-8)(3)BnN +] [FeCl3Br-])MIL可获得较小的单链和双链DNA的高提取效率,而1,12-di( 3-十六烷基苯并咪唑鎓)十二烷双[((三氟甲基)磺酰基]酰亚胺溴三氯高铁酸盐(III)([(C16BnIM)(2)C-122(+)] [NTf2-,FeCl3Br-])MIL对于较大的DNA分子产生更高的提取效率。基于MIL的方法还用于从含有白蛋白的复杂基质中提取DNA,揭示了四氯高铁酸三己基(十四烷基)phosph([P-6,6,6,14(+)与[(C-8)(BnN +)-Bn-3] [FeCl3Br-] MIL相比,] [FeCl4-])MIL的结果明显降低了白蛋白的共提取。采用MIL-DNA方法从细菌细胞裂解物中提取质粒DNA。从MIL提取阶段回收了具有足够质量和数量可用于聚合酶链反应(PCR)扩增的DNA,这证明了在进行下游分析之前,基于MIL的DNA样品制备的可行性。

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