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

Kevin D. Clark, Matthew Sorensen, Omprakash Nacham, Jared L. Anderson

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)39846-39851
Number of pages6
JournalRSC Advances
Volume6
Issue number46
DOIs
StatePublished - 2016

Bibliographical note

Funding Information:
The authors thank the Chemical Measurement and Imaging Program at the National Science Foundation (Grant no. CHE-1413199) for funding this research.

Publisher Copyright:
© The Royal Society of Chemistry 2016.

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