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基于非硅纳米颗粒的核酸分离方法的开发及实时 PCR 试剂盒在食用油溯源中的应用。

Development of nucleic acid isolation by non-silica-based nanoparticles and real-time PCR kit for edible vegetable oil traceability.

机构信息

College of Sciences, Henan Agricultural University, Zhengzhou 450002, China; Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, Zhengzhou 450002, China.

College of Life Sciences, Henan Agricultural University, Zhengzhou 450002, China; Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, Zhengzhou 450002, China.

出版信息

Food Chem. 2019 Dec 1;300:125205. doi: 10.1016/j.foodchem.2019.125205. Epub 2019 Jul 18.

Abstract

For efficient extraction of amplifiable DNA from edible vegetable oils, we developed a novel DNA extraction approach based on the non-silica-based dipolar nanocomposites. The nanoparticle comprises a hydrophilic polymethyl methacrylate core with abundant capillaries, hydrophilic vesicles decorated with molecules having DNA affinity and a coating hydrophobic polystyrene layer. The nanoparticles are soluble in oil, adsorb the DNA from the aqueous phase and gave a high DNA recovery ratio. All DNA extracts from fully refined vegetable oil soybean, peanut, rapeseed, and cottonseed oils, including their blends, were sufficiently pure to be amplified by real-time PCR targeting the chloroplast ribulose-1,5-bisphosphate gene (rbcL), therefore, the species of origin and their ratios in mixed vegetable oils blended from two or three oil-species could be determined. These results indicate that the novel DNA isolation and real-time PCR kit is a simple, sensitive and efficient tool for the species identification and traceability in refined vegetable oils.

摘要

为了从食用植物油中高效提取可扩增 DNA,我们开发了一种基于非硅基偶极纳米复合材料的新型 DNA 提取方法。该纳米颗粒由具有丰富毛细管的亲水性聚甲基丙烯酸甲酯核、带有 DNA 亲和力分子的亲水囊泡和涂覆有疏水性聚苯乙烯层组成。纳米颗粒可溶于油中,从水相中吸附 DNA,并获得高 DNA 回收率。从全精炼植物油(大豆、花生、油菜籽和棉籽油)及其混合物中提取的所有 DNA 均足够纯净,可通过实时 PCR 扩增叶绿体核酮糖-1,5-二磷酸基因(rbcL),因此可以确定混合植物油中起源物种及其比例。这些结果表明,新型 DNA 分离和实时 PCR 试剂盒是一种简单、灵敏、高效的工具,可用于鉴定精炼植物油中的物种和可追溯性。

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