Qualitative and quantitative detection of genetically modified products is inseparable from the application of reference materials (RMs). In this study, a batch of genomic DNA (gDNA) certified reference materials (CRMs) was developed using genetically modified rice Kemingdao (KMD) homozygotes as the raw material. The gDNA CRMs in this batch showed good homogeneity; the minimum sample intake was determined to be 2 μL. The stability study showed that transportation by cold chain is preferable, no significant degradation trend was observed during a 12-month period when storing the gDNA CRMs at 4 °C and − 20 °C, and the number of freeze-thaw cycles cannot exceed 10. The property values of the copy number ratio of transgene and endogenous gene and the copy number concentration for gDNA CRMs were determined by a collaborative characterization of eight laboratories using the duplex KMD/PLD droplet digital PCR (ddPCR) assays. The uncertainty components of characterization, potential between-unit heterogeneity, and potential degradation during long-term storage were combined to estimate the expanded uncertainty of the certified value with a coverage factor k of 2.0. The certified value of copy number ratio for KMD gDNA CRM is 0.99 ± 0.05, and that of copy number concentration is (1.76 ± 0.10) × 10⁵ copies/μL. Compared to the gDNA CRMs in availability, this batch of KMD gDNA CRMs is assigned accurate property values and can be directly used for qualitative and quantitative detection of GMOs as well as evaluation of the parameters of analytical methods with no need of further DNA concentration measurement.

转基因产品的定性定量检测离不开标准物质（RMs）的应用。本研究以转基因水稻克明道（KMD）纯合子为原料，开发了一批基因组DNA（gDNA）认证参考物质（CRMs）。该批次中的 gDNA CRM 显示出良好的均质性；最低样品摄入量确定为 2 μL。稳定性研究表明，冷链运输更可取，gDNA CRMs在4°C和-20°C下储存12个月期间没有观察到明显的降解趋势，并且冻融循环次数不能超过10 .PLD液滴数字 PCR (ddPCR) 检测。将表征的不确定性分量、潜在的单元间异质性和长期储存期间的潜在退化相结合，以估计覆盖系数k为 2.0的认证值的扩展不确定性。KMD gDNA CRM的拷贝数比认证值为0.99±0.05，拷贝数浓度为（1.76±0.10）×10 ⁵拷贝/μL。与现有的gDNA CRM相比，这批KMD gDNA CRM具有准确的属性值，可直接用于GMO的定性和定量检测以及分析方法参数的评估，无需进一步测量DNA浓度。