Ionic liquid–based aqueous biphasic system as an alternative tool for enhanced bacterial DNA extraction

Abstract

BackgroundDeveloping an alternative and benign method for DNA extraction is imperative due to the high cost and potential harms associated with conventional techniques. Investigation of Ionic liquid (IL) as a solvent for DNA storage and stability revealed the ability of IL to assist DNA processes. IL–based aqueous biphasic system emerges as a comprehensive extraction platform capitalizing on the task–specificity of ILs and the wide applicability of ABS for biomolecule extractions. Therefore, it is beneficial to optimize an IL–based ABS specifically for DNA extraction, taking into account the fundamental interactions between the IL and DNA. ResultsThe primary objective was to design ABS consisting of Ammonium based ILs, and Potassium phosphate buffer as the salting–out agent for the partitioning of salmon sperm DNA. The analysis focused on optimizing biocompatible anions for the extraction. Moreover, the stability of the DNA in the IL rich phases was analysed to validate the method. The proposed process was then employed for extracting plasmid DNA from bacteria, demonstrating results comparable to those obtained with a commercially available kit. Further validation using agarose gel electrophoresis and transformation of the extracted DNA into E.coli were conducted, producing promising outcomes. Although there is room for improvement in terms of recovery of DNA and reusability of ABS, the described approach is comparable with the conventional one while being cost–effective, and showcases a noticeable and convincing link to eco–friendly processes. SignificanceThere is limited literature on IL–based ABS for DNA extraction, and the existing studies predominantly concentrate on systems derived from Cholinium ILs. However, their high hydrophilicity limits the choice of the second–phase forming component to polymers for the formation of ABS. Ammonium ILs efficiently form biphasic systems with various available salting–out agents, and biocompatible anions are introduced to mitigate the toxicity of the ILs.