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水热法制备磁性微球及其在植物基因组DNA提取分离中的应用
引用本文:杜鲜,孙培培,张虹,张富昌,石峰.水热法制备磁性微球及其在植物基因组DNA提取分离中的应用[J].安徽农业科学,2016,44(13).
作者姓名:杜鲜  孙培培  张虹  张富昌  石峰
作者单位:石河子大学生命科学学院,新疆石河子,832000;石河子大学生命科学学院,新疆石河子,832000;石河子大学生命科学学院,新疆石河子,832000;石河子大学生命科学学院,新疆石河子,832000;石河子大学生命科学学院,新疆石河子,832000
基金项目:石河子大学高层次人次科研启动项目(RCZX201138),石河子大学科学技术研究发展计划项目(2012ZRKYQ-YD14)。
摘    要:目的]建立一种适用于多种植物组织快速提取基因组DNA的方法。方法]以水热反应制备出Fe3 O4磁性纳米微球,并对其粒径、形貌、磁学性质等进行表征分析,并以此作为核酸提取载体,对多种新疆特色经济作物和植物的叶片、根、茎、籽粒等组织进行DNA的提取分离,并优化分离纯化过程的各环节。结果]通过OD260紫外吸收值的检测、琼脂糖凝胶电泳,结果显示,用该方法纯化得到的植物基因组DNA纯度高、完整性好,能够满足下游分子生物学操作对基因组DNA质量的要求。结论]建立了一种简便、快速、普适的从多种植物组织中获得高产量和高纯度的基因组DNA,为全自动化、规模化提取DNA提供了理论和实践基础。

关 键 词:磁性纳米微球  水热法  基因组DNA  纯化  植物组织

Preparation of Magnetic Nano-microsphere by Solvothermal Synthesis and Its Application in Genomic DNA Purification from Plant Tissues
Abstract: Objective] The aim was to establish a method for rapidly extracting genomic DNA from several kinds of plant tissues. Method] Fe3 O4 magnetic nano-microsphere was prepared by solvothermal synthesis, the surface features of particle size, morphology, magnetic proper-ties were analyzed. With Fe3 O4 magnetic nano-microsphere as nucleic acid extraction vector, DNA of leaves, roots, stems, grains and other tissues of several kinds of Xinjiang special economic crops and plants were extracted and seperated;links of purification and seperation process were optimized. Result] Through detection of OD260 UV absorption value, agarose gel electrophoresis, the results showed that the obtained genomic DNA with high purity and integrity can meet the requirements of the downstream molecular biology operation. Conclusion] A simple and rapid method for obtaining high yield and high quality genomic DNA from several kinds of plant tissues is established, which will provide theoretical and practical basis for automation and large-scale extraction of DNA.
Keywords:Magnetic nano-microsphere  Solvothermal synthesis  Genomic DNA  Purification  Plant tissues
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