Abstract
The efficiency of DNA purification is of particular importance in polymerase chain reaction (PCR)-based diagnostic applications where the quantity of bacterial cells in a potato tissue may be limited. In this work, we report a systematic study of magnetic nanoparticles (MNPs) as probes for genomic DNA isolation from Ralstonia solanacearum and infested potato tubers lysate in the presence of polyethylene glycol (PEG)/NaCl and RNAse A. MNPs were formed under hydrothermal conditions. Particle size diameter and size distribution were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The extracted DNA was tested for its digestibility with two restriction enzymes. PCR-based methods were applied to detect R. solanacearum in potato tuber extracts. The average TEM diameter of MNPs was also approximately 82 nm. MNPs with diameters ranging from 100 to 201 nm were prepared. Herein magnetic nanoparticles provide a reasonable high adsorption amount for DNA. Furthermore, the isolated DNA was found to be compatible in restriction endonuclease digestion and PCR amplification. The high quality of DNA extracted using the modified method was confirmed by the successful amplification of the OLI1/ Y2 specific primers. The magnetic isolation of DNAs offer many benefits, including high quality, simple treatment, quick processing, reduced chemical need and being environment-friendly compared with those obtained by using commercial kits and the boiling method.