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首页> 外文期刊>RSC Advances >Preservation of DNA in nuclease-rich samples using magnetic ionic liquids
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Preservation of DNA in nuclease-rich samples using magnetic ionic liquids

机译：使用磁离子液体保存核酸酶样品中的DNA

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Nucleic acids are important diagnostic molecules for a variety of applications, but are exceedingly sensitive to enzymatic degradation by nucleases. Very recently, hydrophobic magnetic ionic liquids (MILs) have shown considerable promise in the area of DNA extraction. Here, we show that MILs can also serve as DNA preservation media in nuclease-rich environments. DNA samples treated with deoxyribonuclease I (DNase I) were found to retain their molecular weight for up to 72 h at room temperature within the benzyltrioctylammonium bromotrichloroferrate( III ) ([N _(888Bn) ~(+) ][FeCl _(3) Br ~(?) ]) and trihexyl(tetradecyl)phosphonium tetrachloroferrate( III ) ([P _(66614) ~(+) ][FeCl _(4) ~(?) ]) MILs, whereas DNA in aqueous samples suffered complete enzymatic degradation under similar conditions. Using a single drop extraction (SDE) technique, DNase I was found to partition between aqueous solution and MIL with a smaller amount of the enzyme extracted by the [N _(888Bn) ~(+) ][FeCl _(3) Br ~(?) ] MIL relative to the [P _(66614) ~(+) ][FeCl _(4) ~(?) ] MIL. Plasmid DNA (pDNA) exhibited structural stability for up to 1 week in the [N _(888Bn) ~(+) ][FeCl _(3) Br ~(?) ] and [P _(66614) ~(+) ][FeCl _(4) ~(?) ] MILs, even when treated with 20 U of DNase I. pDNA stored within the MIL solvent under these conditions was successfully amplified by polymerase chain reaction (PCR), whereas pDNA in aqueous solutions of DNase I yielded no detectable amplicon. Furthermore, pDNA stored within the trihexyl(tetradecyl)phosphonium tetrachloromanganate( II ) ([P _(66614) ~(+) ] _(2) [MnCl _(4) ~(2?) ]) MIL was capable of conveying antibiotic resistance to competent E. coli following 24 h incubation with DNase I at room temperature, demonstrating that the biological activity of pDNA was preserved.
机译：核酸是各种应用的重要诊断分子，但对核酸酶的酶促降解非常敏感。最近，疏水性磁离子液体（MIL）在DNA提取区域中显示了相当大的希望。在这里，我们表明MILS也可以作为富含核酸酶的环境中的DNA保存介质。发现用脱氧核酸核酸酶I（DNA酶I）处理的DNA样品在苄基三氧基溴氯甲酸酯（III）内的室温下保持其分子量高达72小时（[N _（888Bn）〜（+）] [FECL _（3）四甲基〜（四十四烯）四氯甲酸酯（III）（[P _（66614）〜（+）] [FECL _（4）〜（α）]）米尔，而水性样品中的DNA遭受完整在类似条件下的酶促降解。使用单滴提取（SDE）技术，发现DNase I在水溶液和MIL之间分配，其中[N _（888bn）〜（+）] [feCl _（3）br〜中提取较少量的酶。 （？）]米隆相对于[P _（66614）〜（+）] [FECL _（4）〜（？）] MIL。质粒DNA（PDNA）在[N _（888BN）〜（+）] [FECL _（3）BR〜（α）]和[P _（66614）〜（+）]中表现出长达1周的结构稳定性。 [FECL _（4）〜（α）]米尔，即使用20 u的DNase I.储存在MIL溶剂中的PDNA，通过聚合酶链式反应（PCR）成功扩增，而DNase的水溶液中的PDNA我没有检测到的扩增子。此外，将储存在三氧基（四癸基）四氯蒽烷（II）中的pDNA（[P _（66614）〜（+）] _（2）[MnCl _（4）〜（2？）] mil能够传达抗生素在室温下与DNA酶I孵育24小时后对富集大肠杆菌的抵抗力，证明了PDNA的生物活性。

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