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一种用于检测19个目标以鉴定转基因生物的定性实时PCR方法的开发。

Development of a qualitative real-time PCR method to detect 19 targets for identification of genetically modified organisms.

作者信息

Peng Cheng, Wang Pengfei, Xu Xiaoli, Wang Xiaofu, Wei Wei, Chen Xiaoyun, Xu Junfeng

机构信息

Institute of Quality and Standard for Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021 China ; State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Hangzhou, 310021 China.

College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310058 China.

出版信息

Springerplus. 2016 Jun 24;5(1):889. doi: 10.1186/s40064-016-2395-y. eCollection 2016.

Abstract

As the amount of commercially available genetically modified organisms (GMOs) grows recent years, the diversity of target sequences for molecular detection techniques are eagerly needed. Considered as the gold standard for GMO analysis, the real-time PCR technology was optimized to produce a high-throughput GMO screening method. With this method we can detect 19 transgenic targets. The specificity of the assays was demonstrated to be 100 % by the specific amplification of DNA derived from reference material from 20 genetically modified crops and 4 non modified crops. Furthermore, most assays showed a very sensitive detection, reaching the limit of ten copies. The 19 assays are the most frequently used genetic elements present in GM crops and theoretically enable the screening of the known GMO described in Chinese markets. Easy to use, fast and cost efficient, this method approach fits the purpose of GMO testing laboratories.

摘要

近年来,随着市售转基因生物(GMO)数量的增加,迫切需要分子检测技术的靶标序列具有多样性。作为转基因生物分析的金标准,实时荧光定量PCR技术经过优化,形成了一种高通量转基因生物筛选方法。通过这种方法,我们可以检测19种转基因靶标。通过对来自20种转基因作物和4种非转基因作物的参考材料DNA进行特异性扩增,证明该检测方法的特异性为100%。此外,大多数检测方法显示出非常灵敏的检测效果,检测限达到十个拷贝。这19种检测方法针对转基因作物中最常用的基因元件,理论上能够对中国市场上描述的已知转基因生物进行筛选。这种方法易于使用、快速且成本效益高,适合转基因生物检测实验室的需求。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/999b/4920734/86f83405832c/40064_2016_2395_Fig1_HTML.jpg

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