Rapid method for the detection of Ralstonia solanacerum by isolation DNA from infested potato tubers based on magnetic nanotools
2016
Alghuthaymi, M.A. | Ali, A.A. | Hashim, A.F. | Abd-Elsalam, A.
The efficiency of DNA purification of particular importance in polymerase chain reaction (PCR)-based diagnostics applications where the quantity of bacterial cells in a potato tissue may be limited. In this work, the authors report a systematic study of magnetic nanoparticles (MNPs) a probes for genomic DNA isolation from Ralstonia solanacearum and infested potato tubers lysate into the presence of polyethylene glycol (PEG)/NaCl and RNAse A. MNPs were formed under hypothermal conditions. Particle size diameter and size distribution were characterized by transmission electron microscopy 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 responsible 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 amplifications 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.
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