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Issue 46, 2016, Issue in Progress
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Preservation of DNA in nuclease-rich samples using magnetic ionic liquids

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Abstract

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) ([N888Bn+][FeCl3Br]) and trihexyl(tetradecyl)phosphonium tetrachloroferrate(III) ([P66614+][FeCl4]) 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 [N888Bn+][FeCl3Br] MIL relative to the [P66614+][FeCl4] MIL. Plasmid DNA (pDNA) exhibited structural stability for up to 1 week in the [N888Bn+][FeCl3Br] and [P66614+][FeCl4] 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) ([P66614+]2[MnCl42−]) 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.

Graphical abstract: Preservation of DNA in nuclease-rich samples using magnetic ionic liquids

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Publication details

The article was received on 06 Mar 2016, accepted on 11 Apr 2016 and first published on 13 Apr 2016


Article type: Paper
DOI: 10.1039/C6RA05932E
Citation: RSC Adv., 2016,6, 39846-39851
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    Preservation of DNA in nuclease-rich samples using magnetic ionic liquids

    K. D. Clark, M. Sorensen, O. Nacham and J. L. Anderson, RSC Adv., 2016, 6, 39846
    DOI: 10.1039/C6RA05932E

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