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磁性离子液体作为RNA提取和保存的溶剂

Magnetic Ionic Liquids as Solvents for RNA Extraction and Preservation.

作者信息

Zhu Chenghui, Varona Marcelino, Anderson Jared L

机构信息

Department of Chemistry, Iowa State University, Ames, Iowa 50011, United States.

出版信息

ACS Omega. 2020 May 5;5(19):11151-11159. doi: 10.1021/acsomega.0c01098. eCollection 2020 May 19.

Abstract

Ribonucleic acid (RNA) is particularly sensitive to enzymatic degradation by endonucleases prior to sample analysis. In-field preservation has been a challenge for RNA sample preparation. Very recently, hydrophobic magnetic ionic liquids (MIL) have shown significant promise in the area of RNA extraction. In this study, MILs were synthesized and employed as solvents for the extraction and preservation of RNA in aqueous solution. RNA samples obtained from yeast cells were extracted and preserved by the trihexyl(tetradecyl) phosphonium tris(hexafluoroacetylaceto)cobaltate(II) ([P ][Co(hfacac) ]) and trihexyl(tetradecyl) phosphonium tris(phenyltrifluoroacetylaceto)cobaltate(II) ([P ][Co(Phtfacac) ]) MIL with a dispersion of the supporting media, polypropylene glycol, at room temperature for up to a 7 and 15 day period, respectively. High-quality RNA treated with ribonuclease A (RNase A) was recovered from the tetra(1-octylimidazole)cobaltate(II) di(l-glutamate) ([Co(OIM) ][Glu]) and tetra(1-octylimidazole)cobaltate(II) di(l-aspartate) ([Co(OIM) ][Asp]) MILs after a 24 h period at room temperature. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and agarose gel electrophoresis were used to determine the effect of RNA preservation. Furthermore, the preservation mechanism was investigated by exploring the partitioning of RNase A into the MIL using high-performance liquid chromatography.

摘要

核糖核酸(RNA)在样品分析前对核酸内切酶的酶促降解特别敏感。现场保存一直是RNA样品制备面临的挑战。最近,疏水性磁性离子液体(MIL)在RNA提取领域显示出巨大的潜力。在本研究中,合成了MIL,并将其用作从水溶液中提取和保存RNA的溶剂。从酵母细胞中获得的RNA样品分别在室温下用三己基(十四烷基)鏻双(六氟乙酰丙酮)钴(II)([P ][Co(hfacac) ])和三己基(十四烷基)鏻双(苯基三氟乙酰丙酮)钴(II)([P ][Co(Phtfacac) ])MIL与支撑介质聚丙二醇分散保存长达7天和15天。在室温下放置24小时后,从双(L-谷氨酸)四(1-辛基咪唑)钴(II)([Co(OIM) ][Glu])和双(L-天冬氨酸)四(1-辛基咪唑)钴(II)([Co(OIM) ][Asp])MIL中回收了用核糖核酸酶A(RNase A)处理过的高质量RNA。采用定量逆转录聚合酶链反应(qRT-PCR)和琼脂糖凝胶电泳来确定RNA保存的效果。此外,通过使用高效液相色谱法探索RNase A在MIL中的分配情况来研究保存机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d60f/7241037/661a72e41220/ao0c01098_0001.jpg

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