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Simultaneous cell lysis and DNA extraction from whole blood using magnetic ionic liquids

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Abstract

Conventional DNA sample preparation methods involve tedious sample handling steps that require numerous inhibitors of the polymerase chain reaction (PCR) and instrumentation to implement. These disadvantages limit the applicability of conventional cell lysis and DNA extraction methods in high-throughput applications, particularly in forensics and clinical laboratories. To overcome these drawbacks, a series of nine hydrophobic magnetic ionic liquids (MILs) previously shown to preconcentrate DNA were explored as cell lysis reagents. The MILs were found to lyse white blood cells from whole blood, 2-fold diluted blood, and dry blood samples while simultaneously extracting human genomic DNA. The identity of metal ion incorporated within the MIL appears to cause hemolysis while the cationic component further reduces the cell’s integrity. Over 500 pg of human genomic DNA was isolated from 50 μL of whole blood using the trioctylbenzylammonium tris(hexafluoroacetylaceto)nickelate(II) ([N8,8,8,Bz+][Ni(hfacac)3−]) MIL, and 800 pg DNA was isolated from a dry blood samples using the trihexyl(tetradecyl)phosphonium tris(phenyltrifluoroacetylaceto)nickelate(II) ([P6,6,6,14+][Ni(Phfacac)3−]) MIL following a 1-min vortex step. A rapid, one-step cell lysis and DNA extraction from blood is ideal for settings that seek high-throughput analysis while minimizing the potential for contamination.

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Acknowledgments

M. N. E. would like to thank Paul M. Emaus for his support. J. L. A. acknowledges funding from the Chemical Measurement and Imaging Program at the National Science Foundation (CHE-1709372).

Funding

This study was funded by the Chemical Measurement and Imaging Program at the National Science Foundation (CHE-1709372).

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  1. Department of Chemistry, Iowa State University, 1605 Gilman Hall, Ames, IA, 50011, USA

    Miranda N. Emaus & Jared L. Anderson

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  1. Miranda N. Emaus
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Correspondence to Jared L. Anderson.

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This article contains studies involving human blood samples that were purchased commercially. All studies and procedures have been approved by the Institutional Biosafety Committee (IBC) at Iowa State University.

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The authors declare that they have no conflicts of interest.

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Emaus, M.N., Anderson, J.L. Simultaneous cell lysis and DNA extraction from whole blood using magnetic ionic liquids. Anal Bioanal Chem 412, 8039–8049 (2020). https://doi.org/10.1007/s00216-020-02941-w

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  • DOI: https://doi.org/10.1007/s00216-020-02941-w

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