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.