Detection of genetically modified crops using multiplex asymmetric polymerase chain reaction and asymmetric hyperbranched rolling circle amplification coupled with reverse dot blot

文献类型： 外文期刊

第一作者： Wang, Xiumin

作者： Wang, Xiumin;Teng, Da;Guan, Qingfeng;Wang, Jianhua;Wang, Xiumin;Teng, Da;Guan, Qingfeng;Wang, Jianhua;Tian, Fang

作者机构：

关键词： Hyperbranched rolling circle amplification;Reverse dot blot;Padlock probe;Genetically modified crops

期刊名称：FOOD CHEMISTRY （ 影响因子：7.514； 五年影响因子：7.516 ）

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收录情况： SCI

摘要： To meet the ever-increasing demand for detection of genetically modified crops (GMCs), low-cost, high-throughput and high-accuracy detection assays are needed. The new multiplex asymmetric polymerase chain reaction and asymmetric hyper-branched rolling circle amplification coupled with reverse dot blot (RDB) systems were developed to detect GMCs. Thirteen oligonucleotide probes were designed to identify endogenous targets (Led, Hmg and Sad1), event-specific targets (RRS-5C, RRS-3C, Bt176-3C and MON810-3C), screening targets (35S promoter and NOS terminator), and control targets (18S and PLX). Optimised conditions were as follows: tailed hybridization probes (1-2 pmol/l) were immobilized on a membrane by baking for 2 h, and a 10:1 ratio of forward to reverse primers was used. The detection limits were 0.1 mu g/l of 2% RRS and 0.5 ng/l of DNA from genetically modified (GM) soybean. These results indicate that the RDB assay could be used to detect multiplex target genes of GMCs rapidly and inexpensively. (C) 2014 Elsevier Ltd. All rights reserved.

分类号： TS2`TS201.2

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