Investigation and optimization of DNA isolation efficiency using ferrite-based magnetic nanoparticles

Abstract

DNA isolation is a crucial step in many molecular biological applications for diagnostic and research purposes, like detection of infectious diseases or gene expression studies. However, due to the requirement of toxic reagents in traditional procedures and the high expenses of commercial kits, the use of magnetic MNP-based DNA isolation is becoming more widespread. In this study, different ferrite containing MNPs (MnFe₂O₄, MnFe₂O₄-NH₂, MgFe₂O₄, MgFe₂O₄-NH₂ NiFe₂O₄, NiFe₂O₄-NH₂) are examined and compared in their pDNA isolation efficiency. Among the tested nanoparticles, we document the use of NiFe₂O₄ and its amine-functionalized form for the first time. Three protocols for the isolation of pDNA are optimized for each type of nanoparticle and the best protocol is selected based on the quantity, quality and integrity of the extracted DNA. Plasmid samples extracted with the MNPs are transformed into competent bacterial cells and further tests are performed to recover genomic DNA from bacterial cells, leading to the development of another protocol. Bacteria-spiked blood serum samples are produced to extract DNA from a more complex biological matrix.