生物技术通报 ›› 2013, Vol. 0 ›› Issue (1): 96-101.

• 研究报告 • 上一篇    下一篇

转基因水稻华恢1 号检测用质粒标准分子研制

李亮 ,王晶, 臧超 ,隋志伟 ,赵正宜   

  1. 中国计量科学研究院,北京 100013
  • 收稿日期:2012-04-19 修回日期:2013-01-31 出版日期:2013-01-30 发布日期:2013-01-30
  • 作者简介:李亮,男,博士,助理研究员,研究方向:生物计量学;E-mail: liliangene@yahoo.com.cn
  • 基金资助:

    国家科技支撑计划项目课题(2008BAK41B01)

Development of a Reference Plasmid for the Quantitative Detection of Genetically Modified Rice Huahui-1

Li Liang ,Wang Jing ,Zang Chao ,Sui Zhiwei ,Zhao Zhengyi   

  1. National Institute of Metrology,Beijing 100013
  • Received:2012-04-19 Revised:2013-01-31 Published:2013-01-30 Online:2013-01-30

摘要:

质粒标准分子作为阳性标准品已广泛用于转基因作物及产品的核酸定量检测。但是目前多数质粒标准分子只用于研究,未能成为国家级标准物质。因此,按照国家一级标准物质技术规范,研制了用于检测转基因水稻华恢1 号的质粒标准分子,内容包括质粒分子的构建、纯度分析、特异性检测、可替代性研究、均匀性检验、稳定性考察、定值与不确定度评估。结果表明,该质粒分子标准物质半年内稳定,可替代基因组作为华恢1 号定量的阳性标准。

关键词: 华恢1 , 号 , 转基因水稻 , 实时荧光定量PCR , 标准分子

Abstract:

With the development of genetically modified organism (GMO) detection techniques, the reference plasmids have been themost effective and important materials for GMO quantification as a positive control. But many of them are used in researching, not developeda certified reference material. Here, we outline a reference plasmid, developed according the Technical Norm of Primary Refernce Material(JJF 1006-1994), including preparation, analysis of purify, homogeneity, stability, commutability, characterisation and uncertainty. Resultssuggested the reference molecule is stable for 6 months and particularly suitable for the rapid identification of GMO components in unknownsamples and can reduce cross contamination.

Key words: Huahui 1 , Genetically modified rice , Quantitative real-time PCR , Reference plasmid