Simultaneous Cell Lysis and DNA Extraction using Magnetic Ionic Liquids

Monday, March 08, 2021: 11:05 AM - 11:25 AM

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Description

DNA sample preparation is often times an overlooked bottleneck in DNA analysis. Conventional cell lysis and DNA extraction methods such as detergent lysis methods, silica-based extractions, and cetyltrimethylammonium bromide (CTAB)-based extractions require multiple, time-consuming sample preparation steps that limit their applicability. In addition, conventional methods often require additional DNA purification steps to remove buffers and organic solvents required to lyse cells and extract DNA. To overcome many of the aforementioned disadvantages, a series of magnetic ionic liquids (MILs) were investigated as cell lysis reagents and DNA extraction solvents. MILs are a subclass of ionic liquids that contain a paramagnetic component allowing them to respond to a magnetic field. By combining cell lysis and DNA extraction into a single step, the total DNA sample preparation step could take as short as 1 min, whereas conventional methods take at least an hour. MILs were briefly dispersed in a cell suspension and collected using a rod magnet. The DNA-enriched MIL was then added directly to a custom-buffer for quantitative polymerase chain reaction (qPCR) allowing DNA to desorb during qPCR amplification. The cation, ligand, and chelated metal of the MIL solvents were optimized to detect the largest amount of pure DNA from a cell suspension. The MILs were found to be successful at lysing white blood cells from undiluted blood and from plant cells derived from Arabidopsis thaliana leaves while simultaneously extracting human and plant genomic DNA in minutes allowing for high throughput analysis.

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Additional Info

Keywords: Please select up to 4 keywords ONLY:
Extractions,Nucleic Acids,Clinical Analysis,Sample Preparation