Characterization of iron oxide magnetic nanoparticles for extraction of harar coffee (coffea arabica L.) genomic DNA
摘要
DNA extraction using magnetic nanoparticles (MNPs) has gained significant consideration due to their simple manipulation, cost-effectiveness, and less time-consuming. Even though magnetic separation has been extensively studied for different biological samples, there is a need for efficient methods to extract DNA from woody plants. The present study was aimed to synthesize iron oxide (Fe3O4) MNPs coated with Salicylic Acid (SA) for genomic DNA (gDNA) extraction from Harar coffee leaves. The Fe3O4 MNPs were synthesized from available iron salts under alkaline conditions followed by coating with SA. The synthesized material was characterized by ultraviolet-visible (UV-Vis), X-Ray Diffraction (XRD), Fourier-Transform Infrared (FTIR) spectroscopy, and Scanning Electron Microscopy (SEM). Point of zero charges were determined using the pH drift method. The synthesized Iron oxide magnetic nanoparticles coated with salicylic acid (Fe3O4/SA MNPs) were used for gDNA extraction from coffee leaves. The result of FTIR analysis indicated that, the carboxylic functional group was found in Fe3O4/SA MNPs that indicates successful surface modification. XRD analysis showed that the crystallite phase size of Fe3O4 and Fe3O4/SA MNPs with 14.35 and 10.44 nm respectively. The extraction results revealed that the quality and quantity of extracted gDNA using Fe3O4/SA MNPs were higher than that of the uncoated Fe3O4 and CTAB methods. The PCR assay indicated the amplification of extracted gDNA. The synthesized MNPs indicated respective values of 80% and 67% adsorption and desorption efficiency. The obtained results revealed the effectiveness and convenience of the synthesized Fe3O4/SA MNPs in gDNA extraction from plant leaf samples.