Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 412, Issue 29, Pages 8039-8049Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-020-02941-w
Keywords
Nucleic acids (DNA/RNA); Extraction (SFE/SPE/SPME); White blood cell analysis; Cell lysis; Dry blood analysis
Funding
- Chemical Measurement and Imaging Program at the National Science Foundation [CHE-1709372]
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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 mu L of whole blood using the trioctylbenzylammonium tris(hexafluoroacetylaceto)nickelate(II) ([N-8,8,8,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) ([P-6,6,6,14(+)][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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