Simultaneous cell lysis and DNA extraction from whole blood using magnetic ionic liquids
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Simultaneous cell lysis and DNA extraction from whole blood using magnetic ionic liquids Miranda N. Emaus 1 & Jared L. Anderson 1 Received: 29 July 2020 / Revised: 27 August 2020 / Accepted: 3 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
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) ([N 8 , 8 , 8 , B z + ][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 1min 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. Keywords Nucleic acids (DNA/RNA) . Extraction (SFE/SPE/SPME) . White blood cell analysis . Cell lysis . Dry blood analysis
Introduction Genomic DNA analysis from blood samples is highly important in forensic and clinical applications. Nucleic acid (NA) testing protocols for human immunodeficiency virus (HIV) and hepatitis B virus (HBV) target human genomic DNA, as the virus integrates genomic information into the host’s white blood cells (WBCs) [1–3]. These NA tests are capable of detecting the virus prior to antibody formation and are necessary to rapidly screen blood and organ donations. However, according to the American Red Cross, conventional NA tests for viruses require greater technical skill and expensive equipment compared to antibody testing. In addition, the isolation Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00216-020-02941-w) contains supplementary material, which is available to authorized users. * Jared L. Anderson [email protected] 1
Department of Chemistry, Iowa State University, 1605 Gilman Hall, Ames, IA 50011, USA
of human genomic DNA is common during investigations to determine the source of a bloodstain. In both forensic and clinical applications, there is a great need to rapidly analyze DNA from blood. Sample preparation is often considered an overlooked bottle
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