Optimization of Genomic DNA Extraction with Silica Hydroxyl Magnetic Beads

doi:10.13560/j.cnki.biotech.bull.1985.2016-1045

Abstract

Abstract: This study aims to prepare silicon hydroxyl functionalized magnetic nano magnetic beads and nucleic acid extraction reagents,and to optimize the extraction process. Solvent thermal method was employed to prepare the silicon hydroxyl functionalized nano magnetic beads,then coating SiO₂ on the surface of magnetic beads by surface chemical modification method. The effects of buffer concentration,lysate concentration,dose of magnetic beads and elution temperature on the efficiency of DNA extraction were analyzed by self-made magnetic beads extraction kit,respectively. All the experimental results,quantitatively by ultraviolet spectrophotometer,and qualitatively by agarose gel electrophoresis,were validated;meanwhile,the extraction results were compared with commercial kit. Results showed that we successfully synthesized the Fe₃O₄@SiO₂ magnetic beads of approximate 200 nm. The extraction efficiency of nucleic acid achieved the best on the conditions of magnetic beads was 1.5 mg,lysate contained 5 mol/L guanidine hydrochloride buffer（pH4.9）,the buffer system was 60 mmol/L guanidine hydrochloride（pH7.5）with 60% ethanol,and elution temperature was 65℃. Moreover,there were statistical differences on the effects by the four factors（P < 0.05）. In conclusion,using self-made magnetic beads and reagent system may efficiently carry out the complete genomic DNA extraction of whole blood,even it is better than the commercial kit.

Key words: magnetic nanoparticle, surface modification, genomic DNA extraction

LI Hai-yang, WANG Fei, LEI Hong-tao, ZHANG Xuan, CHEN Ke. Optimization of Genomic DNA Extraction with Silica Hydroxyl Magnetic Beads[J]. Biotechnology Bulletin, 2017, 33(6): 223-229.

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