Separation And Purification Of Genomic DNA From Maize Seeds With Zinc Oxide Nanoparticles

Zinc oxide is a kind of inorganic semiconductor material.Zinc oxide nanoparticles(ZnO NPs)have good biocompatibility,adsorption activity,unique optical and electrical properties.They have shown considerable advantages in catalysis,sensing,drug delivery,bioimaging and many other fields.In this work,ZnO NPs were prepared by using homogeneous precipitation method.The obtained ZnO NPs were characterized by means of UV-vis,TEM,XRD and Zeta potential.ZnO NPs were used for the isolation and purification of DNA,which broadened the application of ZnO NPs in the field of life sciences.The performances of the obtained ZnO nanoparticles as an adsorbent in the isolation and purification of DNA were investigated.A sorption efficiency of 100%was achieved by using 5.0 mg of ZnO nanoparticles for adsorbing 50 ?g mL-1 DNA in 500 ?L sample solution at pH 3.0.The adsorption equilibrium of DNA onto ZnO NPs could be reached within 2 min.The adsorption kinetics was consistent with pseudo-second-order model,and the adsorption rate was mainly controlled by chemisorption.The adsorption behaviors of DNA onto the ZnO NPs fit with Langmuir adsorption model well,corresponding to a theoretical sorption capacity of 60.97 mg g-1.Afterwards,the retained DNA could be readily stripped by using citric acid-sodium citrate buffer,giving rise to a recovery of ca.81.7%.The practical application of the proposed separation and purification protocol was demonstrated by the selective isolation of the genomic DNA from maize seeds.Successful polymerase chain reaction amplification of 88 bp maize ZSSIIb gene was achieved by using the harvested DNA as the template.The results indicated that the DNA extracted from the complex biological sample matrix could meet the requirements of subsequent downstream processes,such as polymerase chain reaction.There was no notable difference in terms of purity and yield between the DNA isolated with the ZnO NPs and that isolated with the plant genomic DNA extraction kit,and the proposed method was very simple without having to strip maize embryos in advance and free from being disturbed by a large number of polysaccharides.